Myofilaments: Movers and Rulers of the Sarcomere

Brian Leei Lin,1 Taejeong Song,1,2 and Sakthivel Sadayappan*1,2

ABSTRACT
Striated cardiac and skeletal muscles play very different roles in the body, but they are similar
at the molecular level. In particular, contraction, regardless of the type of muscle, is a precise
and complex process involving the integral protein myofilaments and their associated regulatory
components. The smallest functional unit of muscle contraction is the sarcomere. Within the sar-
comere can be found a sophisticated ensemble of proteins associated with the thick filaments
(myosin, myosin binding protein-C, titin, and obscurin) and thin myofilaments (actin, troponin,
tropomyosin, nebulin, and nebulette). These parallel thick and thin filaments slide across one an-
other, pulling the two ends of the sarcomere together to regulate contraction. More specifically, the
regulation of both timing and force of contraction is accomplished through an intricate network of
intra- and interfilament interactions belonging to each myofilament. This review introduces the sar-
comere proteins involved in striated muscle contraction and places greater emphasis on the more
recently identified and less well-characterized myofilaments: cardiac myosin binding protein-C,
titin, nebulin, and obscurin. © 2017 American Physiological Society. Compr Physiol 7:675-692,
2017.

Didactic Synopsis Introduction

Major teaching points
1. The sarcomere is the functional unit of muscle contraction
at the molecular level.
2. The sarcomere consists of two sets of myofilaments, which
are made up of rope-like proteins, called myofilament-
associated proteins:
a. Thick filament-associated proteins include myosin,
myosin-binding protein-C, titin, and obscurin.
b. Thin filament-associated proteins include actin, tro-
ponin, tropomyosin, and nebulin.
3. Some myofilaments physically move to affect muscle
contraction:
a. Troponin moves tropomyosin on actin to open a myosin-
binding site.
b. Myosin binds actin and pulls the entire thin filament.
c. Myosin-binding protein-C regulates myosin binding to
actin.
4. While other myofilaments are rulers that measure out the
filament structure, determining where filament proteins are
located on the filament:
a. Titin is the thick filament ruler.
b. Nebulin is the thin filament ruler.
c. Obscurin offers structural support.
5. Many myofilaments have both structural and functional
roles that are still being studied.
The first muscle protein, termed myosin, was discovered and
isolated in 1864 by the German physiologist, Wilhelm Kühne
(131). Subsequent work by Vladimir Engelhardt and Militza
Lyubimova in the late 1930s and 1940s demonstrated that
myosin had enzymatic activity capable of hydrolyzing adeno-
sine triphosphate (ATP) (53). This ATPase activity was found
to drive the interaction of myosin with the second major mus-
cle protein, actin (250). In the 1950s, Andrew Huxley with
coworker Rolf Niedergerke and Hugh Huxley with his col-
league Jean Hanson established the “sliding filament theory”
(103, 105, 106), which holds that muscle contraction is based
on the interaction of myosin and actin filaments. Other ele-
ments of myofilaments, such as tropomyosin (12), troponin
(51), and myosin-binding protein-c (MyBP-C) (199), were
demonstrated around the same time, or shortly thereafter, and
several “giant” sarcomere proteins, including titin (161), neb-
ulin (272), and obscurin (291), were subsequently discovered.
The work by these and other pioneers has laid the foundation
for our understanding of striated muscle function and the
mechanisms controlling contraction.
Cardiac muscles are responsible for circulating the blood
throughout the body, which is primarily performed by the
* Correspondence to sadayasl@ucmail.uc.edu
1 Department of Cell and Molecular Physiology, Health Sciences
Division, Loyola University Chicago, Maywood, Illinois, USA
2 Department of Internal Medicine, Heart, Lung and Vascular Institute,
Division of Cardiovascular Health and Sciences, University of
Cincinnati College of Medicine, Cincinnati, Ohio, USA
Published online, April 2017 (comprehensivephysiology.com)
DOI: 10.1002/cphy.c160026
Copyright © American Physiological Society.

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Myofilament Proteins in Myocyte Contraction
blood-pumping action of the left ventricle (61, 84). Cardiac
muscles are under the control of the autonomic nervous sys-
tem (233). Individual cardiac muscle cells, or cardiomyocytes,
are binucleated and are linked via intercalated disks to con-
tract as a syncytium (244). Contraction occurs via a process
known as excitation-contraction coupling (19), in which an
electrical stimulus is linked to mechanical contraction move-
ment via calcium-induced calcium release (54). Briefly, an
electrical signal, which is modulated in part by autonomic
innervation, travels via the transverse tubules, or T-tubules,
resulting in an initial inward flux of Ca2+ from L-type voltage-
dependent Ca2+ channels known as dihydropyridine receptors
(DHPR). This initial Ca2+ release event is detected by ryan-
odine receptors (RyR2 in the heart) that release Ca2+ stores
from the sarcoplasmic reticulum (SR) (55, 80), thus initiating
contraction. In contrast, skeletal muscle performs a differ-
ent set of functions, such as postural support and voluntary
movement, correspondingly orchestrated by the somatic ner-
vous system. Skeletal muscle is comprised of different fiber
types, and a single fiber can stretch from tendon to tendon-
constituting mechanotransduction of force (242). Similar to
cardiac myocytes, skeletal muscle fibers are multinucleated,
having hundreds of peripheral nuclei that dot the fiber from
one end to another (99, 202). Contraction of individual skele-
tal muscle fibers is dependent on the innervation of each fiber
(102). Activation of one subset of muscle fibers innervated
by one motor neuron may not affect activation of adjacent
muscle fibers. Unlike cardiomyocytes, the electrical signal
is transduced via neuromuscular junctions such that RyR
is physically coupled to DHPR. In addition, after injury,
skeletal muscles have regenerative capability arising from a
reserve of muscle-specific stem cells (186,290). Despite these
differences between cardiac and skeletal muscles, the under-
lying functional unit of contraction, known as the sarcomere,
is very similar. The present review will focus on the contractile
elements of the sarcomere in cardiac muscle.
The Sarcomere
The sarcomere is the smallest unit of striated muscle contrac-
tion and is delineated by Z-disks, the dark striations that differ-
entiate these muscles from smooth muscle (Fig. 1) (122,292).
The sarcomere is situated between two Z-disks, and the M-
line, the attachment site for thick filaments, is located at the
center of the sarcomere. The sarcomere consists of highly
organized components that interact in a very precise manner
to effect contraction at the molecular level (104, 105). The
primary components of contraction are a series of rope-like
proteins known as myofilaments consisting of thick and thin
filaments that run parallel to one another and perpendicular
to the Z-disks. The thin and thick filaments extend from the
Z-disk and M-line, respectively, overlapping on each side of
the M-line and creating distinctive regions within the sarcom-
ere. Under electron microscopy, the light region immediately
adjacent to each Z-disk is known as the I-band (107). A darker
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Comprehensive Physiology
region farther toward the center of the sarcomere is known as
the A-band and is followed by another light region in the cen-
ter known as the H-zone on either side of the M-line. Within
the A-band is a region known as the C-zone consisting of 7 to
9 transverse stripes that run perpendicular to the thick and thin
filaments (42). The various contractile proteins reside within
in these regions, but they can span across these regions as
well. In the sarcomere, locus is a determinant of function in
regulating the initiation of thick and thin filament sliding, the
mechanism underlying contraction.
Thick and thin filaments
The primary component of a thin filament is actin, a heli-
cal rope-like structure attached at the Z-disks (Fig. 1). The
“giant” proteins nebulin and nebulette in skeletal and cardiac
muscles, respectively, span the length of the thin filament.
Along the length of actin, troponin complex proteins period-
ically interact with tropomyosin, which also forms a helical
structure. Myosin, the primary component of thick filaments,
is the motor that drives the sliding activity of actin filaments.
Myosin has several distinct regions, including head, neck,
and tail domains. The “giant” protein titin runs the entire
length of the thick filament, and both obscurin and MyBP-C
are associated with it. By its central location between thick
and thin filaments, MyBP-C is believed to modulate contrac-
tion through its activation/inhibition of the thin filament and
generation of cross-bridge power. A large body of research
has found mutations of key human sarcomeric proteins in
dilated (DCM), hypertrophic (HCM), and restricted (RCM)
cardiomyopathies (Table 1). In this review, we will highlight
aspects of these myofilament proteins by following the gen-
eral order in which they come into play during contraction.
The Troponins
In both cardiac and skeletal muscles, Ca2+ at the sarcomeric
level initiates contraction. The influx of Ca2+ and the signal it
induces are detected and transduced by the troponin complex
that consists of three subunits: Troponin C, Troponin I, and
Troponin T (TnC, TnI, and TnT, respectively) (Fig. 1C). Since
their initial discovery by Setsuro Ebashi (50), much has been
learned regarding the function of each troponin, and such
knowledge has directly led to improved cardiac therapies.
For example, cardiac TnI and TnT are clinically important
biomarkers commonly used to detect and assess the severity
of cardiac injury (10, 197), and mutations in troponins cause
diverse cardiomyopathies (Table 1) (114, 179, 234).
Troponin C
After Ca2+ influx into the sarcomere, the first troponin
engaged is TnC, a calcium-binding 18-kDa troponin sub-
unit within the N-terminal regulatory domains. The isoforms
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(A)
Comprehensive Physiology

I-band A-band I-band

C-zone H-zone C-zone

M-line

Z-disk Z-disk
(B) (C)
Troponin C Tropomyosin
Troponin M
Actin
Nebulette

B
VK
T-Cap

N2
PE
C0 P
Troponin T /A C
1 M
Myosin cMyBP-C
C C2
Myosin
C3

Titin Head
C4

PE
Myosin
C5

Light Chain

VK

N2B
Myosi
y n Tail
C6

α-actinin
C7

Actin Myosin
C8
C9 C10

Figure 1 Structure of sarcomere and myofilament proteins. (A) Actin and myosin overlap at the C-zone in the A-band where 7 to 9 MyBP-C stripes interact with both filaments. Myosin
and MyBP-C are arranged in a ratio of ∼ 3:1. Titin spans from Z-disk to M-line. (B) Titin containing N2B and PEVK regions in I-band anchors in Z-disk via T-cap and α-actinin, and it also
interacts with actin. (C) C-terminal (C8-C10) cMyBP-C binds to meromyosin and titin, but N-terminal C0 to M regions associate with actin and myosin head.

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Table 1 Key Human Sarcomeric Gene Mutations in Cardiomyopathy Troponin I

TnI is a 21-kDa troponin subunit considered to be an important
Cardiomyopathy
Gene Symbol type
inhibitor of sarcomeric activation based on its multiple inter-
actions with other troponin subunits as well as tropomyosin
and actin (200, 294). TnI inhibits actomyosin Mg-ATPase,
Thick filaments
and this inhibition is enhanced by tropomyosin (74, 216). In
β-Cardiac myosin heavy chain MYH7 DCM/HCM/RCM its inactivated state, the N-terminal region of TnI loosely binds
Essential myosin light chain MYL3 HCM TnC and TnT, while its C-terminal region tightly binds actin.
Regulatory myosin light chain MYL2 HCM Both binding affinities and sites of Tnl change upon activation
Cardiac myosin binding protein C MYBPC3 HCM
by Ca2+ . Ca2+ -binding to the N-terminal regulatory domain
of TnC causes C-terminal TnI to tightly bind TnC and TnT,
Obscurin OBSCN DCM
while loosening its interaction with actin (216,254). The bind-
Titin TTN DCM/HCM ing of Ca2+ to TnC causes a conformational change of Tnl
Thin filaments that is vital to its inhibitory effect on actin-activated myosin
α-Cardiac actin ACTC1 DCM/HCM/RCM ATPase and is thought to transduce the signal down to the
last troponin subunit, TnT. TnI has also been demonstrated
Nebulette NEBL DCM/HCM
to be a potent regulator of phosphorylation-mediated Ca2+ -
Cardiac troponin C TNNC1 DCM/HCM sensitivity. Phosphorylation of cTnI by protein kinase A and C
Cardiac troponin I TNNI3 DCM/HCM/RCM (PKA/PKC) decreases force development, but increases car-
Cardiac troponin T TNNT2 DCM/HCM/RCM diac muscle relaxation (117, 194). A loss-of-function study
knocked down the gene expression of cTnl and resulted in
α-Tropomyosin TPM1 HCM
impairment of normal cardiac structure and function (101).
Data Source: www.ncbi.nlm.nih.gov/clinvar, unless otherwise noted.
DCM, dilated cardiomyopathy; HCM, hypertrophic cardiomyopathy; Troponin T
RCM, restricted cardiomyopathy.
At 36 kDa, TnT is the largest of the troponin subunits. It
anchors the other troponin subunits to actin and binds to
tropomyosin (Tm) (7, 94-96). TnT is composed of two heli-
cal structures (T1 and T2) as well as an unstructured flex-
of TnC have somewhat confusing terminology. The cardiac ible region known as the hypervariable domain (256). The
isoform of TnC (cTnC) and slow-skeletal isoform of TnC C-terminal T2 structure interacts with TnI, TnC, and actin
(ssTnC) are structurally identical, being encoded by the same (87, 241) as well as the central region of Tm (187, 210). The
gene. The use of either cTnC or ssTnC refers to the same central T1 region interacts with the C-terminus of Tm. The
protein in cardiac or skeletal muscles, respectively. How- function of the N-terminal hypervariable domains (H1 and
ever, fast-skeletal TnC (fsTnC) is a distinct isoform, varying H2) remains unclear, but evidence suggests that these regions
slightly in primary structure from that of cTnC and ssTnC may allow TnT to adapt to specific isoforms, muscles, or
(71). All TnCs are thought to be similar in structure and species (22, 110). The primary role of TnT is thought to
function, consisting of two globular domains at the N- and be structural, but recent studies suggest that TnT has sev-
C-terminal ends (91, 240) connected via an α-helical bridge eral phosphorylation sites that regulate Ca2+ sensitivity and
(269). The N-terminal globular domain is a primary regula- force generation (109, 195, 251). For example, the interaction
tory region, having low Ca2+ -binding affinity in its two helix- between Tm and TnT, a crucial regulatory step in thin filament
loop-helix (EF-hand) motifs (I and II) (232). C-terminal EF activation (279), is calcium dependent. When Ca2+ binds to
hand motifs (III and IV) bind with high affinity to Mg2+ and TnC, TnT disengages from interaction with Tm, allowing Tm
Ca2+ under physiological conditions, but turnover is too slow to move its position relative to actin such that its myosin-
for the continuation of myofilament activation (294). Thus, binding site is exposed (217).
the C-terminal domain is considered to serve a structural role
by anchoring TnC to the N-terminal domain of TnI (232). In
cTnC, the first EF hand motif (I) is inactivated by substitu-
tions of a key Ca2+ ligand which results in only one functional
Tropomyosin
Ca2+ -binding site at the N-terminus. The N-terminal domain Tropomyosin (Tm) (7) is a 37-kDa protein that is 42 nm in
of TnC interacting with the C-terminal region of TnI has been length. It consists of double α-helices that form a coiled-coil
dubbed the “regulatory head” (121,256). The binding of Ca2+ that intertwines with actin filaments (285,286). Tm consists of
to the N-terminal globular domain partially opens a hydropho- the multiple isoforms α-Tm, β-Tm, and slow α-Tm encoded
bic region of cTnC and alters its interaction with cTnI (246). by TPM1, TPM2, and TPM3, respectively (213). Mutations
The conformational change in the regulatory head transduces in Tm are implicated in both cardiac and skeletal myopathies
the Ca2+ signal. (Table I) (173, 255). These isoforms are structurally similar,

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but variations in structural homology suggest differential
capacity to regulate contraction (190). Tm polymers can form
both homodimers and heterodimers. In striated muscles, Tm
is arranged in heterodimers (108, 148) that overlap by 8 to 9
amino acids from the N-terminus of one Tm polymer to the
C-terminus of another (171). Similar to TnT, one of its binding
partners, Tm is thought to be a stabilizer thin filament, since
interaction between Tm and tropomodulin reduces depoly-
merization of the thin filament components. Functionally, Tm
works in tandem with the troponin complex to regulate thin
filament activation and, hence, myosin-actin interaction. Tm
isoforms dimerize and polymerize end-to-end, fitting into
the major groove within the actin double α-helix (213). In
this groove, Tm physically blocks the myosin-binding site
on actin, thereby preventing contraction by interfering with
strong myosin and actin interaction (59). The position of Tm
in relation to actin directly contributes to the primary func-
tion of Tm in regulating contraction. Tm physically blocks the
myosin-binding sites on actin and thus prevents actin-myosin
binding when the muscle is in its relaxed state. Removal of this
blockage requires a conformation change in Tm and is neces-
sary for the initiation of contraction. The conformational shift
is induced by Ca2+ -bound troponin and causes Tm to shift its
position from blocked to closed position, thus exposing the
myosin-binding sites on actin (170). The binding of myosin to
actin further shifts the position of Tm on actin from closed to
open position, which results in further shifting of the position
of Tm on actin (236).
Actin
Actin is the primary component of the thin filament. Func-
tionally, actin serves to anchor troponins and Tm. In addition,
actin transduces force generation via interaction with thick-
filament proteins, most notably, myosin. The vertebrate actin
gene is complicated, consisting of three highly homologous
isoforms (α, β, and γ) as well as skeletal, cardiac, and smooth
variations of the α isoform (268). Skeletal and cardiac iso-
forms are differentially expressed, and they vary by muscle
type and species. Actin isoforms are highly conserved with
only minimal interspecies variation at the N-terminal region
(268), and differences of just 15 amino acids can result in
major structural and functional variations (30, 31).
Actin is a double α-helical protein that forms a coiled-
coil within the sarcomere (98, 154). Globular actin (G-actin)
polymerizes into filamentous actin (F-actin) in a sponta-
neous manner. G-actin consists of four subdomains (SD1-
SD4) (113,172,243) that form a binding pocket for adenosine
diphosphate (ADP), or ATP (52,119,120) as well as the diva-
lent ions Mg2+ and Ca2+ (204, 205). Seven actin monomers,
comprising SD1/2, are found per half-helical turn of the actin
filament. This defined region interacts with its opposing strand
via SD 3/4 (123). SD 1/2 can also interact with other myofil-
ament proteins, such as myosin, MyBP-C (Fig. 1A) and other
contractile proteins (270). Specifically, actin SD1 binds to the
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Myofilament Proteins in Myocyte Contraction
myosin head, also known as myosin subfragment 1, or myosin
S1, primarily via two known ionic interactions. First, nega-
tively charged SD1 interacts with the positively charged lysine
myosin 50k/20k loop. Second, an adjacent SD1 interacts with
myosin amino acid residues 567 to 578 (175,176,228). A third
interaction occurs between myosin S1 (amino acids 404-415)
and proline residues between actin SD1 and SD3.
Myosin
Myosin is a superfamily of proteins with many classes and
functional roles (58, 127, 287). This review will focus on
class ll myosin in striated muscles, which serves as a cross-
bridge responsible for muscle contraction. As previously
mentioned, myosin ll was originally discovered in 1864 by
Wilhelm Kühne (131). Myosin (∼220 kDa) binds to actin
and, following a conformational change induced by calcium,
“pulls” the thin filaments across the thick filaments. Typical
of many proteins involved in muscle contraction, myosin
itself consists of several distinct regions, including the heavy
meromyosin fragment, which is itself comprised of the
head (S1) and neck (S2), and the light meromyosin (LMM),
comprising the long tail of the protein. The N-terminal region
has a head region known as the S1 subfragment (49,228,229).
The S1 subfragment projects outward from the thick filament
at 14.3-nm intervals and physically interacts with actin to
form cross-bridges. This catalytic head is the motor that drives
sarcomeric muscle contraction (262). The S1 subfragment is
primarily responsible for the sliding action whereby thick and
thin filaments are able to alter their relative parallel position
to each other. In the rigor or bound state, myosin is bound to
actin without an attached nucleotide. The catalytic head also
possesses a nucleotide-binding pocket capable of converting
chemical energy (ATP) into mechanical energy for contrac-
tion (97). The release of inorganic phosphate and ADP causes
the myosin head, which is bound to actin, to shift the relative
positions of the thin/thick filaments in an energy-demanding
power stroke. Upon ATP binding to the myosin head, myosin
detaches from actin, myosin Mg-ATPase activity hydrolyzes
the ATP into ADP and inorganic phosphate, and the myosin
head returns to its original position at which point the cycle
can be repeated (97).
Two major isoforms of myosin ll are present in heart
muscle: α-myosin and ß-myosin expressed by the MYH6
and MYH7 genes, respectively (222, 257). The expression of
myosin isoforms is species-dependent (153, 157, 158, 191),
and mutations of myosin genes are a common cause of
hypertrophic cardiomyopathy (23,39,184). More specifically,
rodent species primarily express the faster α-isoform, while
larger mammals predominantly express the slower ß-isoform
(215). This expression pattern may be altered in heart dis-
ease. In rodent species, myosin transitions from α to ß, with
distinct functional consequences in regards to contractile
velocity at the cellular level and cardiac dysfunction at the
whole organ level (128, 181, 253, 258). Conversely, humans
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Cardiac MyBP-C (cMyBP-C, 140 kDa)

N’ C0 P/A C1 M C2 C3 C4 C5 C6 C7 C8 C9 C10 C’

Fast-skeletal MyBP-C (fsMyBP-C, 128 kDa)

P/A C1 M C2 C3 C4 C5 C6 C7 C8 C9 C10

Slow-skeletal MyBP-C (ssMyBP-C, 129 kDa)

P/A C1 M C2 C3 C4 C5 C6 C7 C8 C9 C10

lg domain

Fn3 domain
M domain
Proline/Alanine rich region
Phosphorylation site

Figure 2 Schematic diagram of MyBP-C isoforms. All three isoforms, cardiac, slow skele-
tal, and fast skeletal, contain a proline/alanine (P/A)-rich region, seven immunoglobulin-
like domains (Ig), M-domain, and three fibronectin 3 (Fn3) domains. Top: cardiac isoform
of MyBP-C has an additional C0 Ig domain at the N-terminus and 28 residual inserts in
C5 (brown vertical line). In addition, five phosphorylation sites (red vertical line) can be
observed, one in the P/A region and four in M-domain. The linker (thick purple band)
between C4 and C5 is conserved between cMyBP-C and ssMyBP-C. Middle: fast skele-
tal MyBP-C isoform is the smallest, and no phosphorylation site has been reported. Bottom:
slow skeletal isoform of MyBP-C has four phosphorylation sites, three in the P/A region and
one in the M-region.

exhibit a transition from approximately 93% ß and 7% α to
almost exclusively ß-myosin during heart failure (178). These
differences become important when considering potential
therapeutic targets and clinical outcomes of potential ther-
apeutic regimens, particularly with respect to genetic defects
(69). Just below the S1 subfragment is the myosin neck, also
known as the S2 subfragment, which links the S1 subfragment
to the rest of the thick filament. In addition, the S1 lever arm
interacts with two additional components, known as myosin
light chains (MLCs) to fine-tune myosin function. These light
chain proteins are known as the essential light chain (ELC)
(16) and regulatory light chain (RLC). The ELC and RLC are
localized well within the myosin S1 and S2 region, and as
such, they are able to regulate myosin and, possibly, MyBP-
C function (247). However, the precise regulatory roles of
ELC and RLC remain to be fully defined. Farther toward the
C-terminus is the LMM. The LMM forms the coiled coil that
anchors myosin to the thick filament where it interacts with
titin and MyBP-C.
Myosin-Binding Protein-C
Interaction between myosin and actin is regulated by
MyBP-C, first relegated to the status of an “impurity” dur-
ing myosin purification in the 1970s (42, 199). Subsequently,
three distinct MyBP-C isoforms were identified, including
slow skeletal, fast skeletal, and cardiac, encoded by the dis-
tinct genes MYBPC1, MYBPC2, and MYBPC3, respectively
(32, 278) (Fig. 2). Mutations in MYBPC3 are now known to
be the most common cause of hypertrophic cardiomyopa-
thy (33). MyBP-C isoforms are localized within the C-zone
(199), a region within the sarcomere flanking the M-line (70),
and are arranged in 7 to 9 transverse stripes at 43-nm intervals
(17,46,56,235) (Fig. 1A). As the names suggest, slow-skeletal
and fast-skeletal isoforms (ssMyBP-C and fsMyBP-C, respec-
tively) are predominantly expressed in skeletal muscles and
conditionally expressed in the heart, albeit at lower levels
(47, 48, 149). The cardiac isoform (cMyBP-C) is expressed
exclusively in the heart (65). The 43-nm interval of MyBP-C
and 14.3-nm interval of myosin heads correlate to a 1:3 ratio
between MyBP-C and myosin within the C-zone (42, 155).
MyBP-C structure
MyBP-C has seven immunoglobulin (Ig) and three
fibronectin III (Fn3) domains, numbered C1 through C10,
from the N-terminus to the C-terminus (156). In addition, a
proline/alanine-rich (PA) region precedes the C1 Ig region, as
well as an M-motif between C1 and C2 domains (15,68,134).
Despite these similarities in the C-terminus, MyBP-C
isoforms are structurally distinct by virtue of many key
differences within the N-terminal region. Specifically, cardiac
MyBP-C (cMyBP-C) has an additional immunoglobulin-like

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C-terminal Ig domain, C0, an insertion in the C5 domain of
28 amino acids (227), and five known phosphorylation sites,
including four within the M-motif (41,180) and one in the PA
region (136). In contrast, ssMyBP-C has one known phospho-
rylation site in the M-motif and three phosphorylation sites in
the PA region (2) (Fig. 2). Currently, no phosphorylation sites
have been identified in fsMyBP-C, but evidence suggests
the presence of phosphorylation sites based on changes in
the phosphorylation state of both ssMyBP-C and fsMyBP-C
during disease (1, 3, 4).
MyBP-C regulates contraction primarily via protein-
protein interactions, as little evidence has shown either enzy-
matic activity or interaction with nucleic acids and sugars
(214). The C-terminal region of MyBP-C has been demon-
strated to interact with the LMM region of myosin (177, 185,
201) and titin (60, 64) in the thick filament. These domains
(C8-C10) are vital for localization to the A-band of the
sarcomere (70). MyBP-C interaction with titin determines
localization and periodicity of MyBP-C within the sarcomere
(64,126). Titin, with its eleven domain super-repeats (a series
of immunoglobulin and fibronectin III domains), acts as a
scaffold for MyBP-C within the sarcomere (138, 263, 264).
The C-terminal domains of MyBP-C may also participate in
the organization and stability of thick filaments during devel-
opment (196). While C-terminal MyBP-C domains anchor
MyBP-C to the thick filament, MyBP-C N-terminal binding
partners are more varied with correspondingly more complex
functional implications. The cMyBP-C-knockout mouse car-
diomyopathy models reveal that sarcomere structures are still
conserved (83, 168). Normal sarcomere structure is observed
in human patients with MyBP-C mutations that result in
hypertrophic cardiomyopathy and distal arthrogryposis type 1
(257, 271), suggesting that the role of MyBP-C in sarcomeric
assembly is not necessary.
MyBP-C function
Since it was discovered that mutations in MyBP-C are the
most frequent genetic etiology of hypertrophic cardiomyopa-
thy, interest in understanding its structural/functional mecha-
nisms of action has grown. MyBP-C is frequently considered
a braking system in cardiac muscle contraction (218, 231)
based on its ability to reduce thin filament sliding velocities;
consequently, loss-of-function mutations may overwork the
cardiac muscle, leading, in turn, to the gradual onset of heart
failure in susceptible individuals. Furthermore, the N-terminal
region of MyBP-C binds to actin, thereby shifting Tm from
blocked to closed position in a manner similar to that of the
Ca2+ -bound troponin complex (189). Studies using recom-
binant proteins representing the cMyBP-C N-terminus have
demonstrated that cMyBP-C significantly promotes Ca2+ sen-
sitivity and the rate of tension redevelopment (230,281). This
key function of MyBP-C is likely accomplished by a com-
plicated coordinated interaction with actin (245, 248, 280),
myosin subfragment-2 (S2) (77, 78, 185, 199, 249), and RLC
of myosin (227). While the name myosin-binding protein-C
Volume 7, April 2017
Myofilament Proteins in Myocyte Contraction
suggests only association with the thick filament, studies have
clearly demonstrated that MyBP-C also interacts with thin
filament. Both the C1 domain and M-motif are required to
connect with actin (21, 248, 280), and the cardiac-specific
C0 domain of cMyBP-C interacts with actin to elicit some
functional effects (90). In contrast, MyBP-C interaction with
myosin subfragment-2 appears to be limited to the M-motif,
specifically the N-terminal 126 residues of S2 (77). It is not
yet clear whether the interactions of cMyBP-C with actin and
myosin S2 are mutually exclusive (21, 77). Indeed, the litera-
ture is inconsistent on the role of C0 in contraction. Kunst et al.
demonstrated that the C0 domain alone is not sufficient for
interaction with myosin (133), but other investigators demon-
strated that C0 does interact with the RLC of myosin (227).
The discrepancies suggest subtle differences between differ-
ent isoforms, species, and systems used in these experiments.
Studies demonstrating N-terminal MyBP-C binding to
both actin and myosin suggest not only a unique orientation
between the thin and thick filaments but also a promiscuity
with multiple binding partners regulated in a phosphorylation-
dependent manner (15, 238) (Fig. 1C and 2). Importantly,
phosphorylation of cMyBP-C ablates its interaction with
myosin subfragment-S2, resulting in greater force production
by promoting myosin-actin binding (132). Specifically, phos-
phorylation of MyBP-C regulatory sites (S276, S285, and
S304 in humans) predisposes N-terminal binding to actin,
rather than myosin, a shift in binding preference that may
allow cMyBP-C to provide regulation at multiple levels during
contraction. First, MyBP-C interaction with myosin is hypoth-
esized to act as a mechanical load on myosin and, hence,
reduce its ability to form cross-bridges. Second, MyBP-C
phosphorylation reverses this “braking” activity by releasing
the load on myosin and thus promoting cross-bridge cycling
kinetics. Third, cMyBP-C M-motif phosphorylation confers a
specific structural stiffness to the region and results in a con-
formation change in the N-terminal region (40, 219). Upon
phosphorylation, the N-terminal region of cMyBP-C folds
on itself, thereby potentially shielding residues responsible
for myosin binding within the M-motif, freeing N-terminal
MyBP-C to interact with actin and, hence, facilitate contrac-
tion. However, at high Ca2+ concentration, this N-terminal
folding is ablated (207, 219), suggesting that this region of
cMyBP-C would once again favor myosin binding. This
model allows for binding of MyBP-C to both myosin and
actin at different stages of contraction, that is, both low and
high Ca2+ , but whether the binding interactions are mutually
exclusive has yet to be resolved.
Titin
Titin is a sarcomeric thick filament spanning the length of
the half sarcomere; the N-terminal titin anchors at the Z-
disk, and the C-terminal overlaps at the M-line. Titin is the
third myofilament system in the sarcomere. It was the first
of three “giant” myofilament proteins to be discovered (161).
Originally named “connectin” by Koscak Maruyama in 1976
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Myofilament Proteins in Myocyte Contraction Comprehensive Physiology

Titin isoforms
N2B
N’ N2B PEVK C’

P P P

N2BA
N2B N2A PEVK

P P P

Ig domain

N2B unique sequence

PEVK element
P Phosphorylation site

Figure 3 Domain structure in I-band region of two adult titin isoforms. Titin is a giant myofilament
protein consisting of mostly Ig tandem repeats (orange), but also unique regions specific to each
isoform (81). Both isoforms consist of proximal poly-Ig repeats, a variably spliced region, followed
by distal poly-Ig repeats. Isoform differences are notably distinct within the variable region. Top:
N2B notably contains an N2B unique sequence, N2B-U. Bottom: N2BA consists of a PEVK element
within its variable region.

or, alternatively, “titin” by Kuan Wang in 1979, titin was and
still is not only the largest myofilament protein in the sar-
comere (162, 163, 276), but also the largest known protein.
Titin (TTN) is a single gene that codes for 38,138 amino
acids (4.2 MDa) in humans (13). While alternative splic-
ing yields many different splice variants, three major splice
classes of cardiac titin have been identified: N2B, N2BA, and
FCT (Fig. 1B and 3) (13, 143). At approximately 3.0 MDa,
N2B is the stiffest and shortest isoform, and it is the predomi-
nant isoform in rodent left ventricle (∼80%). N2BA is the next
longest isoform at 3.3 to 3.5 MDa, and it is more compliant,
containing an additional element rich in proline (P), glutamic
acid (E), lysine (K), and valine (V) residues referred to as the
PEVK domain and Ig domains. The ratio of N2BA to N2B
varies among species with increasing amounts of N2BA in
larger mammals (35). Both N2B and N2BA titin isoforms are
expressed in adult cardiac muscle. Because N2B is stiffer than
N2BA (50), which results in differential compliance, changes
in the ratio between the two affect passive tension within the
sarcomere.
Not surprisingly, pathologic heart conditions are asso-
ciated with altered titin expression. For example, the N2B
isoform is increased in animal models of DCM and hyper-
tension, potentially accounting for the increased passive
stiffness in affected muscle (277, 288). Alternatively, a
significant increase of N2BA:N2B ratio observed in patients
with heart failure (206) may serve to improve diastolic
function by decreasing muscle stiffness (192). Indeed, titin
mutations in human patients are most commonly associated
with DCM (89), although HCM-associated titin mutations
have been reported (166, 239). Disease-causing mutations
appeared to be common within the A-band region, but some
mutations were also found in N2B and N2BA regions,
suggesting that dysfunction along the length of titin can
result in cardiomyopathies. In addition, N2BA expression is
significantly elevated in patients with coronary artery disease,
possibly a compensatory response to the increasing stiffness
of cardiac muscle compromised by accumulated collagen
(193). Fetal cardiac titin (FCT) is the longest isoform at 3.6 to
3.8 MDa. FCT is expressed exclusively during development
in the fetal isoform and has even more Ig regions and PEVK
repeats (75, 143, 203). Clearly, the variable expression of titin
isoforms with differential compliance may allow the heart to
adapt to different conditions. A more detailed discussion of
structural and functional implications can be found below.
Titin structure
Titin anchors to the Z-disk and extends across the I-band and
A-band toward the M-line. Titin interactions within the Z-disk
are mediated by α-actinin and telethonin (76, 141, 151, 188).
The Z-disk region of titin consists of approximately 2000
amino acids and interacts with other myofilament proteins,
such as nebulin/nebulette, obscurin, and actin (142,151,291).
This Z-disk region of titin is comprised of three subdomains,
each with distinct structural and functional properties (38).
The first subdomain of titin contains two Ig-repeat domains
designated Z1 and Z2. The second subdomain is a phosphory-
latable linker region that is rich in serine and proline residues
(66). The third subdomain contains another Z domain (Z3),
followed by 45-amino acid ‘Z-repeats’ designated Zr (66).

682 Volume 7, April 2017

Comprehensive Physiology
The I-band consists of a combination of tandem Ig
domains, N2B, N2A, and PEVK elements. Among them, Ig
domains and PEVK elements are crucial extensible sites con-
ferring elasticity to the I-band region of titin (283). There
exist two different isoforms of adult cardiac titin (N2B and
N2BA), which are classified by variable splicing of region of
TTN that encodes the I-band region (Fig. 1). Both isoforms
contain a proximal poly-Ig tandem repeat region, followed by
a variable spliced region and a distal poly-Ig tandem repeat
region (152) (Fig. 3). Within the variable region, the N2B
is composed of Ig domains and a unique sequence known as
N2B-U (72). The N2A region consists of Ig repeats, but also a
prominent PEVK region that can interact with both actin and
Ca2+ (37, 137). Of particular interest, I-band titin is highly
phosphorylated by various kinases (5, 8, 82, 289).
Titin within the A-band is composed of repetitive
sequences of the immunoglobulin superfamily, Ig, and the
fibronectin type III superfamily, Fn3, known as “super-
repeats” that consist of six 7-domain repeats toward Z-disk
or eleven repeats of eleven domains. The C-terminus of titin
at the M-line comprises several binding domains, including
serine/threonine kinase domains (67), a binding domain for
muscle-specific calpain p94 (118), four-and-a-half LIM pro-
tein (DRAL/FHL-2) (144), and a muscle-specific ring finger
protein (MURF-1) (36). M-line titin binds the myosin filament
via myomesin at the sarcomere midpoint (63, 267).
Titin function
Titin is best known as a key regulator of passive tension.
Titin is thought of as a “molecular spring” with elastic recoil
properties that contribute to passive tension during diastolic
relaxation (88,152). However, the function of titin is complex,
and this “giant” protein can be divided into distinct regions
with different functions. Within the Z-disk, titin Z1 and Z2
domains interact with telethonin (76, 188) in an antiparallel
fashion that resists high mechanical stress (20, 147). While
the precise mechanism of this resistance remains to be fully
elucidated, experimental alterations to this antiparallel config-
uration results in reduced mechanical stability. Interestingly,
the resistance by titin/telethonin interaction only occurs in
the direction of physiological contraction. In other words, the
structure does not confer resistance to force or stretch perpen-
dicular to the direction of thick and thin filaments. Titin inter-
action with a Z-disk-associated protein, known as α-actinin,
contributes to structural support for the Z-disk and anchors
the remainder of titin protein (38, 76, 292). Z-disk titin may
also participate in mechanosignaling, but the precise mech-
anism has yet to be fully determined (73, 221). Indeed, the
entirety of titin has been implicated as a key player in signal
transduction based on its enormous length and the role of its
elastic regions in regulating passive tension (152). The pri-
mary elements responsible for this elasticity are localized at
the I-band region of titin (265, 266). When the sarcomere is
stretched, the domains that make up the “spring” within the
I-band region also begin to stretch in a specific and ordered
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Myofilament Proteins in Myocyte Contraction
manner. When the sarcomere contracts, titin elongates, with
the N-terminal Ig repeats stretching first, followed by the
PEVK region (150, 274). Further contraction results in the
unfolding of the PEVK region as I-band titin is stretched
(152, 274). Interestingly, the cardiac-specific isoform N2B
element is capable of further structural unfolding upon addi-
tional stretching of the cell (152, 265, 289).
Another important function of titin is to provide a scaffold
for thick filament proteins within the A-band region. The Ig
and Fn3 super-repeats of A-band titin closely associate with
one another to both confer structural stability to this region
and establish a particular periodicity (86). These super-repeats
span approximately 44 nm in length, coinciding with the peri-
odicity of myosin within the A-band and MyBP-C within
the C-zone. Furthermore, the 11 super-repeats, reflecting the
number of transverse stripes, reveal the presence of MyBP-C
contacting titin within the C-zone (60). The disruption of
interactions between MyBP-C C-terminus and titin results
in the mislocalization of MyBP-C and cardiac dysfunction
(135). Based on its periodicity and scaffolding function, titin
has been termed the “thick filament ruler.” Similar to a ruler
that defines regular, periodic intervals, titin appears to mea-
sure out the regular, periodic intervals of interacting proteins.
The C-terminal region of titin also has functional properties.
At the M-line, it binds to several proteins, including p94,
DRAL/FHL-2, and MURF-1 (198, 267). Although the func-
tion of these binding partners have yet to be fully elucidated,
they are thought to serve several roles, such as regulating
myofibril assembly, structural integrity, and signal transduc-
tion (211, 276). Furthermore, like I-band titin, M-line titin
contains phosphorylatable serine/threonine domains (67).
The regulation of passive tension is thought to take place
at the level of variable splicing and posttranslational modifi-
cation of titin. Phosphorylation presents a faster system for
modulating passive tension of titin compared with isoform
turnover. Titin has multiple phosphorylation sites, including
three in the I-band region, and multiple protein kinases tar-
get the same, or different, sites in the I-band, all of which
affect its ability to develop passive force (5, 73, 82, 289). For
example, protein kinase A (PKA) phosphorylates the N2B
element in I-band and reduces passive tension of rodent and
human cardiac muscles in vitro (130, 289). Cyclic guanosine
monophosphate-dependent protein kinase G phosphorylates
the same site with a similar effect (129). However, phos-
phorylation of PEVK element by PKC has the opposite
effect by increasing stiffness of myocardium (92). Titin is
also phosphorylated by extracellular signal-regulated kinase 2
and calcium/calmodulin-dependent protein kinase II. The full
scope of titin function, as regulated by phosphorylation, has
sparked considerable interest (82, 93, 226).
Due to its massive size, many aspects of titin regulation
are still unclear. In addition, regulation of titin by its numer-
ous binding partners is still being elucidated, including RNA-
binding motif protein 20 (Rmb20), T-cap, a Z-disk-associated
protein, and muscle-ring-finger proteins 1/2 (MURF1/2). For
example, Rbm20 induces alternative splicing of PEVK, and
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Myofilament Proteins in Myocyte Contraction
N’
Acidic region Serine-rich region
SH3 domain Nebulin repeat
Nebulin super-repeat
Figure 4 Schematic illustration of nebulin (top, >600 kDa) and nebulette (bottom, 107 kDa)
structures. Nebulin spans the entire length of thin filament and has a central core region. This
central sequence consists of 8 nebulin repeats, 22 nebulin super-repeats, and an additional
23 nebulin repeats. Nebulette, the cardiac counterpart of nebulin, is much shorter and lacks
nebulin super-repeats.
its mutation results in DCM (79, 293). Interactions between
N-terminal Ig domain and T-cap are not crucial for tight
anchoring of titin to the Z-disk (100). However, T-cap does
mediate E3 ubiquitin ligase-induced titin degradation, in con-
junction with mouse-double-minute-2-homolog (260), sug-
gesting that the degradation of titin is highly regulated. Indeed,
other sites along titin are also known to regulate its degra-
dation. This includes the titin kinase domain, a catalytic
region activated by mechanical stress, which was implicated
in the autophagy/lysosomal pathway (167,220). In the M-line
region, three domains, including Ig141/Ig142/FNIII132, con-
nect to MURF1/2 (284), but the function is still unclear. The
contribution of titin to length-dependent activation has also
been demonstrated (45, 165, 259), but its precise mechanism
has only recently begun to be elucidated (6). The enormity
of titin and its interactions throughout the sarcomere underlie
its many multifaceted roles within the sarcomere. Titin’s mul-
tiple interactions with various binding partners, such as the
enigmatic nebulin and obscurin, underscore the difficulty of
fully understanding its contribution to function in health and
disease.
Nebulin/Nebulette
Nebulin (600-900 kDa) and nebulette (107 kDa) are mem-
bers of the nebulin family of actin-binding cytoskeletal
myofilaments that regulate thin filament length. Nebulin is
predominantly expressed in skeletal muscles, while nebulette
is the predominant isoform in cardiac muscles (14, 116).
Other known members include N-RAP, lasp-1, and lasp-2, the
smallest of which is a mere 39 kDa, demonstrating the diver-
sity of nebulin proteins that regulate sarcomeric structure and
function (208).
Nebulin/nebulette structure
Nebulin extends along the length of a thin filament; the
C-terminus binds to Z-line, and the N-terminus stretches
684
Comprehensive Physiology
Nebulin
C’
Nebulette
toward M-line (275). Similarly, the nebulette C-terminus
localizes in the Z-disk (174, 182). Both nebulin and nebulette
share certain structural elements. They both have an acidic
N-terminal region whose structure has yet to be determined.
This is followed by a central region consisting of tandem
repeats of a conserved sequence of SDxxYK amino acids,
termed as nebulin-repeat motif, ending in a C-terminal region
that is a serine-rich and capped by an SH3 domain (139, 140)
(Fig. 4). However, the predominant skeletal isoform (nebulin)
and the cardiac isoform (nebulette) also exhibit differences,
particularly in the central nebulin repeat region. First, in the
central region, nebulin possesses 8 nebulin repeats followed
by 22 nebulin “super-repeats.” In turn, each super-repeat con-
sists of seven nebulin-repeats as well as an additional con-
served motif with amino acids WLKGIGW that occurs after
every three nebulin-repeats within the super-repeat. This is
then followed by an additional 23 nebulin-repeats. The period-
icity of these tandem repeats is important for interactions with
the thin filament proteins actin, troponin, and Tm (140, 273).
In contrast, the central region of nebulette only consists of 23
nebulin-repeats. The smaller size and complete lack of neb-
ulette’s characteristic super-repeat ultrastructure have poten-
tial functional implications other than regulating thin filament
length.
Nebulin/nebulette function
The super-repeat structure of nebulin’s central region has a
specific periodicity that matches that of the thin filament
structure. For example, seven successive nebulin-repeats of
the super-repeat coincide with seven actin monomers per turn.
Interactions between nebulin and actin are particularly strong
(111), and nebulin notably interacts with other thin filament-
associated proteins (112), suggesting a role as a scaffold for
thin filament proteins. These scaffolding interactions con-
trol protein periodicity and thin filament length, and as such,
nebulin, similar to titin, has been described as a “thin fila-
ment ruler” (14, 207, 209, 282). Just as titin interacts with its
Volume 7, April 2017
Comprehensive Physiology
scaffolding partners at seemingly regular intervals, nebulin
also appears to make contact with actin at regular intervals.
However, the function of nebulin as a ruler is less certain.
First, although different knockout mouse models lacking neb-
ulin have demonstrated that skeletal myocytes exhibit shorter
and/or nonuniform thin filaments, they are not completely
ablated (14, 261, 282). This suggests that nebulin may be dis-
pensable for early developmental stages in muscle develop-
ment or that the expression of other components may com-
pensate for the loss of nebulin. Indeed, nebulin appears to be
capable of regulating the minimal thin filament length, but not
its final length (34). Therefore, while nebulin contributes to
the determination of thin filament length by stabilizing the thin
filament, it is not the sole determinant of thin filament, and,
hence, the term “thin filament ruler” may be an oversimplifi-
cation of the role of nebulin and nebulette in the sarcomere.
Finally, only low levels of nebulin have been detected in the
heart. Expression of nebulin was detected in about 50% of
atrial tissue and only within small regions of endocardial ven-
tricular tissue (14). Knockdown of nebulin in cardiomyocytes
causes elongation and restriction of thin filament; however,
unlike skeletal muscle where structural deficiencies are appar-
ent, no abnormal cardiac phenotype develops in vitro (169).
The cardiac counterpart of nebulin, nebulette is much
smaller in comparison (107 kDa), and it is specifically
expressed in cardiac muscle (182). The smaller nebulette
localizes to the Z-disk and is approximately one-sixth of
the length of nebulin. As such, it is unlikely that nebulette
can determine thin filament protein periodicity along the full
length of the thin filament. Instead, nebulette may stabilize
Z-disk structure (164, 182). However, in vitro disruption of
nebulette in cardiomyocytes results in structural and func-
tional deficits, such as loss of Tm and reduced cardiomyocyte
beating frequency (183). These structural deficits are compa-
rable to those of nebulin knockout mice in which thin filament
lengths were reduced (14,261), suggesting that nebulette reg-
ulates thin filament lengths with additional contribution from
other stabilizing proteins (14). Loss of nebulette also reduces
cardiomyocyte function in vitro (24,183), but the direct cause
is unclear. Despite difference in size, these studies agree that
nebulette and nebulin share a similar function in stabilizing
the thin filament. However, in nebulette knockout mice, nei-
ther cardiac dysfunction nor structural misalignments could
be detected with the exception of increased Z-line width (164).
More research is required to elucidate the functional role and
precise mechanisms of action of these two less well-studied
striated muscle proteins.
Obscurin
The most recently identified member of the “giant” myofil-
ament family is obscurin (700-900 kDa), named in 2001 by
Mathias Gautel and colleagues (291). Obscurin is encoded by
the OBSCN gene and is expressed in striated muscles, with
two known isoforms, obscurin A and obscurin B (29). An
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Myofilament Proteins in Myocyte Contraction
additional shorter isoform of obscurin A, “mini-A” obscurin,
is also thought to exist (62). Within the heart, obscurin is more
highly expressed in ventricles relative to atria and is localized
at the Z-disk, A/I junction, and M-line, depending on the iso-
form (27). While much remains to be elucidated regarding
obscurin, recent studies have begun to unravel its structure
and function.
Obscurin structure
Obscurin isoforms arise from alternative splicing and primar-
ily consist of tandem repeats of 49 Ig domains with two dis-
tinct FN3 domains. While obscurin A and B share several con-
served structural elements, obscurin isoforms differ in their
combination of additional C-terminal domain elements, such
as IQ, SH3, Rho-GEF, and PH domains, depending on the
isoform (13,237,252,291). The C-terminus of obscurin A con-
sists of an SH3 domain, Rho-GEF, and PH domain; obscurin
B has an additional IQ and two protein kinase domains that
arise from alternative splicing (Fig. 5). These differences in
structure may allow different obscurin isoforms to localize to
a specific region of the sarcomere. Obscurin A localizes to the
Z-disk and M-line via its interaction with small ankyrin 1, or
sAnk1 (11, 125, 225). Obscurin B localizes to the M-line and
A/I junction (29).
Obscurin function
One of the first distinguishing properties of obscurin at the
time of its discovery was its interaction with the Z9 and Z10
domains of titin, an interaction that localizes obscurin to titin
at the Z-disk (13, 291). These Z-disk interactions contribute
to structural support in the sarcomere during development
(25, 291). Knockdown of obscurin disrupts the M-line and
A-band regions, but not the Z-disk region (124, 224). At the
M-line, obscurin interacts with the C-terminus of titin (M10
domain) and myomesin (63). Obscurin C-terminal regions are
associated with ankyrin proteins at cardiac M-line, and the
C-terminal regions are necessary for targeting ankyrin
proteins to the M-line (44). The three primary isoforms of
ankyrin are ankyrin 1, ankyrin 2, and ankyrin 3, also known as
ankyrin R, ankyrin B, and ankyrin G, respectively (18,43,85).
However, obscurin has been demonstrated to interact with
only muscle-specific ankyrin 1 and 2 (44,125,291). Obscurin
interactions with ankyrin at the M-line are thought to be
responsible for the lateral alignment of myofilaments, yet
neither structural nor functional deficits are evident in
obscurin-knockout mice (145). This is complicated by the
presence of an obscurin-like 1 protein (Obsl1) that also inter-
acts with myomesin and titin (63). The presence of Obsl1 may
therefore compensate for the lack of obscurin during sarcom-
eric development. Future studies will define whether Obsl1
is an obscurin isoform or a different protein altogether. How-
ever, further analysis of obscurin-deficient mice demonstrates
that the lack of obscurin destabilizes ankyrin proteins, par-
ticularly sAnk1 (145, 146). As obscurin is thought to anchor
685
Myofilament Proteins in Myocyte Contraction
Obscurin A
N’
Obscurin B
Ig domain IQ domain
Fn3 domain SH3 domain
Figure 5 Domain patterns of obscurin A and B. Both isoforms are homologous, consisting
of Ig tandem repeats, Fn3 and IQ domains. The C-terminal domains of obscurin isoforms
are more variable with, for example, additional protein kinase domains and Ig domains.
sAnk1 to the M-line, mislocalization of sAnk1 throughout the
sarcomere results in structural changes to the SR and central-
ized nuclei. Different interactions among various isoforms of
obscurin and ankyrin may regulate contractility. The impor-
tance of these variations in the structure-function relationship
is underscored by the changes in obscurin isoform expression
in cardiac disorders, such as DCM, and obscurin mutations
(10,25,152,153,274). Further studies are required to elucidate
the function of obscurin associated with cardiac pathology
(9, 26, 159, 160, 288). Like all sarcomere proteins discussed
above, a thorough understanding of obscurin’s function is by
no means complete since closer examination of regulatory
domains suggests the presence of various kinase and GEF
domains known to have roles in cell adhesion (100) and cell
signaling, such as the RhoA pathway (28, 57, 212, 223).
Conclusion
In the present review, we focused on the major myofilament
proteins within the context of their role in cardiac muscle
contraction. Over the past century or so, different experimen-
tal methods have been used to discover these myofilaments
and determine their functional roles. Mutations in the genes
encoding myofilament proteins highlight their functional
importance. Even a single mutation can disrupt the intricate
myofilament machinery, underscoring the complexity of their
interactions with one another and with other elements in the
sarcomere. However, beyond characterizing the impact of
discrete genetic defects, much more needs to be learned about
their function in the sarcomere. For example, myofilaments
also have important signaling roles within the muscle
architecture, regulating Ca2+ -sensitivity, length-dependent
activation, gene expression, and cellular stress responses. As
686
Comprehensive Physiology
C’
RhoGEF domain Protein kinase domain
PH domain
research methods continue to improve, the nature of the
dynamic and extraordinarily complicated interplay among
myofilament proteins will be better understood. Moving for-
ward, the expanded understanding of myofilament structure,
function and regulation will undoubtedly broaden the scope of
treatment options available for muscle diseases.
Acknowledgements
The authors were supported by National Institutes of Health
grants R01HL130356, R01HL105826, and K02HL114749
(S. Sadayappan), the American Heart Association Midwest
Affiliate Research Programs (Cardiovascular Genome-
Phenome Study (15CVGPSD27020012), and Grant-in-Aid
(14GRNT20490025).
References
1. Ackermann MA, Kerr JP, King B, C WW, Kontrogianni-
Konstantopoulos A. The phosphorylation profile of myosin binding
protein-C slow is dynamically regulated in slow-twitch muscles in
health and disease. Sci Rep 5: 12637, 2015.
2. Ackermann MA, Kontrogianni-Konstantopoulos A. Myosin binding
protein-C slow is a novel substrate for protein kinase A (PKA) and C
(PKC) in skeletal muscle. J Proteome Res 10: 4547-4555, 2011.
3. Ackermann MA, Patel PD, Valenti J, Takagi Y, Homsher E, Sellers
JR, Kontrogianni-Konstantopoulos A. Loss of actomyosin regulation in
distal arthrogryposis myopathy due to mutant myosin binding protein-C
slow. FASEB J 27: 3217-3228, 2013.
4. Ackermann MA, Ward CW, Gurnett C, Kontrogianni-Konstantopoulos
A. Myosin binding protein-C slow phosphorylation is altered in
Duchenne dystrophy and arthrogryposis myopathy in fast-twitch skele-
tal muscles. Sci Rep 5: 13235, 2015.
5. Ahmed SH, Lindsey ML. Titin phosphorylation: Myocardial passive
stiffness regulated by the intracellular giant. Circ Res 105: 611-613,
2009.
6. Ait-Mou Y, Hsu K, Farman GP, Kumar M, Greaser ML, Irving TC, de
Tombe PP. Titin strain contributes to the Frank-Starling law of the heart
Volume 7, April 2017
Comprehensive Physiology
by structural rearrangements of both thin- and thick-filament proteins.
Proc Natl Acad Sci U S A 113: 2306-2311, 2016.
7. Alves ML, Dias FA, Gaffin RD, Simon JN, Montminy EM, Biesiadecki
BJ, Hinken AC, Warren CM, Utter MS, Davis RT, III, Sadayappan
S, Robbins J, Wieczorek DF, Solaro RJ, Wolska BM. Desensitization
of myofilaments to Ca2+ as a therapeutic target for hypertrophic car-
diomyopathy with mutations in thin filament proteins. Circ Cardiovasc
Genet 7: 132-143, 2014.
8. Anderson BR, Bogomolovas J, Labeit S, Granzier H. The effects of
PKCalpha phosphorylation on the extensibility of titin’s PEVK element.
J Struct Biol 170: 270-277, 2010.
9. Arimura T, Matsumoto Y, Okazaki O, Hayashi T, Takahashi M, Inagaki
N, Hinohara K, Ashizawa N, Yano K, Kimura A. Structural analysis of
obscurin gene in hypertrophic cardiomyopathy. Biochem Biophys Res
Commun 362: 281-287, 2007.
10. Babuin L, Jaffe AS. Troponin: The biomarker of choice for the detection
of cardiac injury. CMAJ 173: 1191-1202, 2005.
11. Bagnato P, Barone V, Giacomello E, Rossi D, Sorrentino V. Binding
of an ankyrin-1 isoform to obscurin suggests a molecular link between
the sarcoplasmic reticulum and myofibrils in striated muscles. J Cell
Biol 160: 245-253, 2003.
12. Bailey K. Tropomyosin: A new asymmetric protein component of mus-
cle. Nature 157: 368, 1946.
13. Bang ML, Centner T, Fornoff F, Geach AJ, Gotthardt M, McNabb M,
Witt CC, Labeit D, Gregorio CC, Granzier H, Labeit S. The complete
gene sequence of titin, expression of an unusual approximately 700-
kDa titin isoform, and its interaction with obscurin identify a novel
Z-line to I-band linking system. Circ Res 89: 1065-1072, 2001.
14. Bang ML, Li X, Littlefield R, Bremner S, Thor A, Knowlton KU, Lieber
RL, Chen J. Nebulin-deficient mice exhibit shorter thin filament lengths
and reduced contractile function in skeletal muscle. J Cell Biol 173:
905-916, 2006.
15. Barefield D, Sadayappan S. Phosphorylation and function of cardiac
myosin binding protein-C in health and disease. J Mol Cell Cardiol 48:
866-875, 2010.
16. Bayram Y, Karaca E, Coban Akdemir Z, Yilmaz EO, Tayfun GA, Aydin
H, Torun D, Bozdogan ST, Gezdirici A, Isikay S, Atik MM, Gambin
T, Harel T, El-Hattab AW, Charng WL, Pehlivan D, Jhangiani SN,
Muzny DM, Karaman A, Celik T, Yuregir OO, Yildirim T, Bayhan IA,
Boerwinkle E, Gibbs RA, Elcioglu N, Tuysuz B, Lupski JR. Molecular
etiology of arthrogryposis in multiple families of mostly Turkish origin.
J Clin Invest 126: 762-778, 2016.
17. Bennett P, Craig R, Starr R, Offer G. The ultrastructural location of
C-protein, X-protein and H-protein in rabbit muscle. J Muscle Res Cell
Motil 7: 550-567, 1986.
18. Bennett V, Chen L. Ankyrins and cellular targeting of diverse mem-
brane proteins to physiological sites. Curr Opin Cell Biol 13: 61-67,
2001.
19. Bers DM. Cardiac excitation-contraction coupling. Nature 415: 198-
205, 2002.
20. Bertz M, Wilmanns M, Rief M. The titin-telethonin complex is a
directed, superstable molecular bond in the muscle Z-disk. Proc Natl
Acad Sci U S A 106: 13307-133310, 2009.
21. Bhuiyan MS, Gulick J, Osinska H, Gupta M, Robbins J. Determination
of the critical residues responsible for cardiac myosin binding protein
C’s interactions. J Mol Cell Cardiol 53: 838-847, 2012.
22. Biesiadecki BJ, Chong SM, Nosek TM, Jin JP. Troponin T core struc-
ture and the regulatory NH2-terminal variable region. Biochemistry 46:
1368-1379, 2007.
23. Bonne G, Carrier L, Richard P, Hainque B, Schwartz K. Familial hyper-
trophic cardiomyopathy: From mutations to functional defects. Circ Res
83: 580-593, 1998.
24. Bonzo JR, Norris AA, Esham M, Moncman CL. The nebulette repeat
domain is necessary for proper maintenance of tropomyosin with the
cardiac sarcomere. Exp Cell Res 314: 3519-3530, 2008.
25. Borisov AB, Kontrogianni-Konstantopoulos A, Bloch RJ, Westfall MV,
Russell MW. Dynamics of obscurin localization during differentia-
tion and remodeling of cardiac myocytes: Obscurin as an integrator of
myofibrillar structure. J Histochem Cytochem 52: 1117-1127, 2004.
26. Borisov AB, Raeker MO, Kontrogianni-Konstantopoulos A, Yang K,
Kurnit DM, Bloch RJ, Russell MW. Rapid response of cardiac obscurin
gene cluster to aortic stenosis: Differential activation of Rho-GEF and
MLCK and involvement in hypertrophic growth. Biochem Biophys Res
Commun 310: 910-918, 2003.
27. Borisov AB, Raeker MO, Russell MW. Developmental expression and
differential cellular localization of obscurin and obscurin-associated
kinase in cardiac muscle cells. J Cell Biochem 103: 1621-1635,
2008.
28. Bowman AL, Catino DH, Strong JC, Randall WR, Kontrogianni-
Konstantopoulos A, Bloch RJ. The rho-guanine nucleotide exchange
factor domain of obscurin regulates assembly of titin at the Z-disk
through interactions with Ran binding protein 9. Mol Biol Cell 19:
3782-3792, 2008.
Volume 7, April 2017
Myofilament Proteins in Myocyte Contraction
29. Bowman AL, Kontrogianni-Konstantopoulos A, Hirsch SS, Geisler
SB, Gonzalez-Serratos H, Russell MW, Bloch RJ. Different obscurin
isoforms localize to distinct sites at sarcomeres. FEBS Lett 581: 1549-
1554, 2007.
30. Brault V, Reedy MC, Sauder U, Kammerer RA, Aebi U, Schoenen-
berger C. Substitution of flight muscle-specific actin by human (beta)-
cytoplasmic actin in the indirect flight muscle of Drosophila. J Cell Sci
112(Pt 21): 3627-3639, 1999.
31. Brault V, Sauder U, Reedy MC, Aebi U, Schoenenberger CA. Dif-
ferential epitope tagging of actin in transformed Drosophila produces
distinct effects on myofibril assembly and function of the indirect flight
muscle. Mol Biol Cell 10: 135-149, 1999.
32. Carrier L, Hengstenberg C, Beckmann JS, Guicheney P, Dufour C,
Bercovici J, Dausse E, Berebbi-Bertrand I, Wisnewsky C, Pulvenis D,
et al. Mapping of a novel gene for familial hypertrophic cardiomyopathy
to chromosome 11. Nat Genet 4: 311-313, 1993.
33. Carrier L, Mearini G, Stathopoulou K, Cuello F. Cardiac myosin-
binding protein C (MYBPC3) in cardiac pathophysiology. Gene 573:
188-197, 2015.
34. Castillo A, Nowak R, Littlefield KP, Fowler VM, Littlefield RS. A
nebulin ruler does not dictate thin filament lengths. Biophys J 96: 1856-
1865, 2009.
35. Cazorla O, Freiburg A, Helmes M, Centner T, McNabb M, Wu Y,
Trombitas K, Labeit S, Granzier H. Differential expression of cardiac
titin isoforms and modulation of cellular stiffness. Circ Res 86: 59-67,
2000.
36. Centner T, Yano J, Kimura E, McElhinny AS, Pelin K, Witt CC,
Bang ML, Trombitas K, Granzier H, Gregorio CC, Sorimachi H,
Labeit S. Identification of muscle specific ring finger proteins as poten-
tial regulators of the titin kinase domain. J Mol Biol 306: 717-726,
2001.
37. Chung CS, Methawasin M, Nelson OL, Radke MH, Hidalgo CG, Got-
thardt M, Granzier HL. Titin based viscosity in ventricular physiology:
An integrative investigation of PEVK-actin interactions. J Mol Cell
Cardiol 51: 428-434, 2011.
38. Clark KA, McElhinny AS, Beckerle MC, Gregorio CC. Striated muscle
cytoarchitecture: An intricate web of form and function. Annu Rev Cell
Dev Biol 18: 637-706, 2002.
39. Colegrave M, Peckham M. Structural implications of beta-cardiac
myosin heavy chain mutations in human disease. Anat Rec (Hoboken)
297: 1670-1680, 2014.
40. Colson BA, Thompson AR, Espinoza-Fonseca LM, Thomas DD. Site-
directed spectroscopy of cardiac myosin-binding protein C reveals
effects of phosphorylation on protein structural dynamics. Proc Natl
Acad Sci U S A 113:3233-3238, 2016.
41. Copeland O, Sadayappan S, Messer AE, Steinen GJ, van der Velden J,
Marston SB. Analysis of cardiac myosin binding protein-C phospho-
rylation in human heart muscle. J Mol Cell Cardiol 49: 1003-1011,
2010.
42. Craig R, Offer G. The location of C-protein in rabbit skeletal muscle.
Proc R Soc Lond B Biol Sci 192: 451-461, 1976.
43. Cunha SR, Mohler PJ. Cardiac ankyrins: Essential components for
development and maintenance of excitable membrane domains in heart.
Cardiovasc Res 71: 22-29, 2006.
44. Cunha SR, Mohler PJ. Obscurin targets ankyrin-B and protein phos-
phatase 2A to the cardiac M-line. J Biol Chem 283: 31968-31980,
2008.
45. de Tombe PP, Mateja RD, Tachampa K, Ait Mou Y, Farman GP, Irving
TC. Myofilament length dependent activation. J Mol Cell Cardiol 48:
851-858, 2010.
46. Dennis JE, Shimizu T, Reinach FC, Fischman DA. Localization of C-
protein isoforms in chicken skeletal muscle: Ultrastructural detection
using monoclonal antibodies. J Cell Biol 98: 1514-1522, 1984.
47. Dhoot GK, Hales MC, Grail BM, Perry SV. The isoforms of C protein
and their distribution in mammalian skeletal muscle. J Muscle Res Cell
Motil 6: 487-505, 1985.
48. Dhoot GK, Perry SV. Expression of slow skeletal myosin binding C-
protein in normal adult mammalian heart. J Muscle Res Cell Motil 26:
143-148, 2005.
49. Dominguez R, Freyzon Y, Trybus KM, Cohen C. Crystal structure of a
vertebrate smooth muscle myosin motor domain and its complex with
the essential light chain: Visualization of the pre-power stroke state.
Cell 94: 559-571, 1998.
50. Ebashi S, Endo M. Calcium ion and muscle contraction. Prog Biophys
Mol Biol 18: 123-183, 1968.
51. Ebashi S, Kodama A. A new protein factor promoting aggregation of
tropomyosin. J Biochem 58: 107-108, 1965.
52. Egelman EH, Orlova A. New insights into actin filament dynamics.
Curr Opin Struct Biol 5: 172-180, 1995.
53. Engelhardt VALMN. Myosin and adenosinetriphosphatase. Nature 144:
668-669, 1939.
54. Fabiato A. Calcium-induced release of calcium from the cardiac sar-
coplasmic reticulum. Am J Physiol 245: C1-C14, 1983.
687
Myofilament Proteins in Myocyte Contraction
55. Fill M, Copello JA. Ryanodine receptor calcium release channels. Phys-
iol Rev 82: 893-922, 2002.
56. Flashman E, Redwood C, Moolman-Smook J, Watkins H. Cardiac
myosin binding protein C: Its role in physiology and disease. Circ
Res 94: 1279-1289, 2004.
57. Ford-Speelman DL, Roche JA, Bowman AL, Bloch RJ. The rho-
guanine nucleotide exchange factor domain of obscurin activates rhoA
signaling in skeletal muscle. Mol Biol Cell 20: 3905-3917, 2009.
58. Foth BJ, Goedecke MC, Soldati D. New insights into myosin evolution
and classification. Proc Natl Acad Sci U S A 103: 3681-3686, 2006.
59. Fraser ID, Marston SB. In vitro motility analysis of actin-tropomyosin
regulation by troponin and calcium. The thin filament is switched as a
single cooperative unit. J Biol Chem 270: 7836-7841, 1995.
60. Freiburg A, Gautel M. A molecular map of the interactions between titin
and myosin-binding protein C. Implications for sarcomeric assembly
in familial hypertrophic cardiomyopathy. Eur J Biochem 235: 317-323,
1996.
61. Friedland G. Discovery of the function of the heart and circulation of
blood. Cardiovasc J Afr 20: 160, 2009.
62. Fukuzawa A, Idowu S, Gautel M. Complete human gene structure of
obscurin: Implications for isoform generation by differential splicing.
J Muscle Res Cell Motil 26: 427-434, 2005.
63. Fukuzawa A, Lange S, Holt M, Vihola A, Carmignac V, Ferreiro A, Udd
B, Gautel M. Interactions with titin and myomesin target obscurin and
obscurin-like 1 to the M-band: Implications for hereditary myopathies.
J Cell Sci 121: 1841-1851, 2008.
64. Furst DO, Vinkemeier U, Weber K. Mammalian skeletal muscle C-
protein: Purification from bovine muscle, binding to titin and the char-
acterization of a full-length human cDNA. J Cell Sci 102(Pt 4): 769-778,
1992.
65. Gautel M, Furst DO, Cocco A, Schiaffino S. Isoform transitions of
the myosin binding protein C family in developing human and mouse
muscles: Lack of isoform transcomplementation in cardiac muscle. Circ
Res 82: 124-129, 1998.
66. Gautel M, Goulding D, Bullard B, Weber K, Furst DO. The central
Z-disk region of titin is assembled from a novel repeat in variable copy
numbers. J Cell Sci 109(Pt 11): 2747-2754, 1996.
67. Gautel M, Leonard K, Labeit S. Phosphorylation of KSP motifs in the
C-terminal region of titin in differentiating myoblasts. EMBO J 12:
3827-3834, 1993.
68. Gautel M, Zuffardi O, Freiburg A, Labeit S. Phosphorylation switches
specific for the cardiac isoform of myosin binding protein-C: A modu-
lator of cardiac contraction? EMBO J 14: 1952-1960, 1995.
69. Geisterfer-Lowrance AA, Kass S, Tanigawa G, Vosberg HP, McKenna
W, Seidman CE, Seidman JG. A molecular basis for familial hyper-
trophic cardiomyopathy: A beta cardiac myosin heavy chain gene mis-
sense mutation. Cell 62: 999-1006, 1990.
70. Gilbert R, Cohen JA, Pardo S, Basu A, Fischman DA. Identification of
the A-band localization domain of myosin binding proteins C and H
(MyBP-C, MyBP-H) in skeletal muscle. J Cell Sci 112(Pt 1): 69-79,
1999.
71. Gomes AV, Potter JD, Szczesna-Cordary D. The role of troponins in
muscle contraction. IUBMB Life 54: 323-333, 2002.
72. Granzier H, Radke M, Royal J, Wu Y, Irving TC, Gotthardt M, Labeit
S. Functional genomics of chicken, mouse, and human titin supports
splice diversity as an important mechanism for regulating biomechanics
of striated muscle. Am J Physiol Regul Integr Comp Physiol 293: R557-
R567, 2007.
73. Granzier HL, Labeit S. The giant protein titin: A major player in
myocardial mechanics, signaling, and disease. Circ Res 94: 284-295,
2004.
74. Greaser ML, Gergely J. Reconstitution of troponin activity from three
protein components. J Biol Chem 246: 4226-4233, 1971.
75. Greaser ML, Krzesinski PR, Warren CM, Kirkpatrick B, Campbell
KS, Moss RL. Developmental changes in rat cardiac titin/connectin:
Transitions in normal animals and in mutants with a delayed pattern of
isoform transition. J Muscle Res Cell Motil 26: 325-332, 2005.
76. Gregorio CC, Trombitas K, Centner T, Kolmerer B, Stier G, Kunke K,
Suzuki K, Obermayr F, Herrmann B, Granzier H, Sorimachi H, Labeit
S. The NH2 terminus of titin spans the Z-disc: Its interaction with a
novel 19-kD ligand (T-cap) is required for sarcomeric integrity. J Cell
Biol 143: 1013-1027, 1998.
77. Gruen M, Gautel M. Mutations in beta-myosin S2 that cause familial
hypertrophic cardiomyopathy (FHC) abolish the interaction with the
regulatory domain of myosin-binding protein-C. J Mol Biol 286: 933-
949, 1999.
78. Gruen M, Prinz H, Gautel M. cAPK-phosphorylation controls the inter-
action of the regulatory domain of cardiac myosin binding protein C
with myosin-S2 in an on-off fashion. FEBS Lett 453: 254-259, 1999.
79. Guo W, Schafer S, Greaser ML, Radke MH, Liss M, Govindarajan
T, Maatz H, Schulz H, Li S, Parrish AM, Dauksaite V, Vakeel P,
Klaassen S, Gerull B, Thierfelder L, Regitz-Zagrosek V, Hacker TA,
Saupe KW, Dec GW, Ellinor PT, MacRae CA, Spallek B, Fischer R,
688
Comprehensive Physiology
Perrot A, Ozcelik C, Saar K, Hubner N, Gotthardt M. RBM20, a gene
for hereditary cardiomyopathy, regulates titin splicing. Nat Med 18:
766-773, 2012.
80. Gurnett CA, Desruisseau DM, McCall K, Choi R, Meyer ZI, Talerico
M, Miller SE, Ju JS, Pestronk A, Connolly AM, Druley TE, Weihl CC,
Dobbs MB. Myosin binding protein C1: A novel gene for autosomal
dominant distal arthrogryposis type 1. Hum Mol Genet 19: 1165-1173,
2010.
81. Ha KM, Cleland H, Greensmith A, Chong D, Macgill K, Verhoeven
A, Hutson JM. Submucous cleft palate: An often-missed diagnosis.
J Craniofac Surg 24: 878-885, 2013.
82. Hamdani N, Krysiak J, Kreusser MM, Neef S, Dos Remedios CG, Maier
LS, Kruger M, Backs J, Linke WA. Crucial role for Ca2(+)/calmodulin-
dependent protein kinase-II in regulating diastolic stress of normal and
failing hearts via titin phosphorylation. Circ Res 112: 664-674, 2013.
83. Harris SP, Bartley CR, Hacker TA, McDonald KS, Douglas PS, Greaser
ML, Powers PA, Moss RL. Hypertrophic cardiomyopathy in cardiac
myosin binding protein-C knockout mice. Circ Res 90: 594-601, 2002.
84. Harvey W. Exercitatio Anatomica de Motu Cordis et Sanguinis in Ani-
malibus. Am J Obstet Gynecol 121: 1007, 1975
85. Hashemi SM, Hund TJ, Mohler PJ. Cardiac ankyrins in health and
disease. J Mol Cell Cardiol 47: 203-209, 2009.
86. Head JG, Houmeida A, Knight PJ, Clarke AR, Trinick J, Brady RL.
Stability and folding rates of domains spanning the large A-band super-
repeat of titin. Biophys J 81: 1570-1579, 2001.
87. Heeley DH, Golosinska K, Smillie LB. The effects of troponin T frag-
ments T1 and T2 on the binding of nonpolymerizable tropomyosin to
F-actin in the presence and absence of troponin I and troponin C. J Biol
Chem 262: 9971-9978, 1987.
88. Helmes M, Trombitas K, Centner T, Kellermayer M, Labeit S, Linke
WA, Granzier H. Mechanically driven contour-length adjustment in rat
cardiac titin’s unique N2B sequence: Titin is an adjustable spring. Circ
Res 84: 1339-1352, 1999.
89. Herman DS, Lam L, Taylor MR, Wang L, Teekakirikul P, Christodoulou
D, Conner L, DePalma SR, McDonough B, Sparks E, Teodorescu DL,
Cirino AL, Banner NR, Pennell DJ, Graw S, Merlo M, Di Lenarda A,
Sinagra G, Bos JM, Ackerman MJ, Mitchell RN, Murry CE, Lakdawala
NK, Ho CY, Barton PJ, Cook SA, Mestroni L, Seidman JG, Seidman
CE. Truncations of titin causing dilated cardiomyopathy. N Engl J Med
366: 619-628, 2012.
90. Herron TJ, Rostkova E, Kunst G, Chaturvedi R, Gautel M, Kentish
JC. Activation of myocardial contraction by the N-terminal domains of
myosin binding protein-C. Circ Res 98: 1290-1298, 2006.
91. Herzberg O, James MN. Refined crystal structure of troponin C from
turkey skeletal muscle at 2.0 A resolution. J Mol Biol 203: 761-779,
1988.
92. Hidalgo C, Hudson B, Bogomolovas J, Zhu Y, Anderson B, Greaser M,
Labeit S, Granzier H. PKC phosphorylation of titin’s PEVK element:
A novel and conserved pathway for modulating myocardial stiffness.
Circ Res 105: 631-638, 617 p following 638, 2009.
93. Hidalgo CG, Chung CS, Saripalli C, Methawasin M, Hutchinson KR,
Tsaprailis G, Labeit S, Mattiazzi A, Granzier HL. The multifunctional
Ca(2+)/calmodulin-dependent protein kinase II delta (CaMKIIdelta)
phosphorylates cardiac titin’s spring elements. J Mol Cell Cardiol 54:
90-97, 2013.
94. Hitchcock SE. Cross-linking of troponin with dimethylimido esters.
Biochemistry 14: 5162-5167, 1975.
95. Hitchcock SE. Regulation of muscle contraction: Bindings of troponin
and its components to actin and tropomyosin. Eur J Biochem 52: 255-
263, 1975.
96. Hitchcock SE, Lutter LC. Study of troponin with cleavable protein
crosslinkers. FEBS Lett 57: 172-174, 1975.
97. Holmes KC, Angert I, Kull FJ, Jahn W, Schroder RR. Electron cryo-
microscopy shows how strong binding of myosin to actin releases
nucleotide. Nature 425: 423-427, 2003.
98. Holmes KC, Popp D, Gebhard W, Kabsch W. Atomic model of the
actin filament. Nature 347: 44-49, 1990.
99. Holtzer H, Marshall JM, Jr., Finck H. An analysis of myogenesis by
the use of fluorescent antimyosin. J Biophys Biochem Cytol 3: 705-724,
1957.
100. Hu LY, Kontrogianni-Konstantopoulos A. The kinase domains of
obscurin interact with intercellular adhesion proteins. FASEB J 27:
2001-2012, 2013.
101. Huang X, Pi Y, Lee KJ, Henkel AS, Gregg RG, Powers PA, Walker
JW. Cardiac troponin I gene knockout: A mouse model of myocardial
troponin I deficiency. Circ Res 84: 1-8, 1999.
102. Hughes BW, Kusner LL, Kaminski HJ. Molecular architecture of the
neuromuscular junction. Muscle Nerve 33: 445-461, 2006.
103. Huxley AF, Niedergerke R. Measurement of muscle striations in stretch
and contraction. J Physiol 124: 46P-47P, 1954.
104. Huxley AF, Niedergerke R. Structural changes in muscle during con-
traction; interference microscopy of living muscle fibres. Nature 173:
971-973, 1954.
Volume 7, April 2017
Comprehensive Physiology
105. Huxley H, Hanson J. Changes in the cross-striations of muscle during
contraction and stretch and their structural interpretation. Nature 173:
973-976, 1954.
106. Huxley HE. X-ray analysis and the problem of muscle. P Roy Soc Lond
B Bio 141: 59-62, 1953.
107. Huxley HE. The double array of filaments in cross-striated muscle.
J Biophys Biochem Cytol 3: 631-648, 1957.
108. Jancso A, Graceffa P. Smooth muscle tropomyosin coiled-coil dimers.
Subunit composition, assembly, and end-to-end interaction. J Biol
Chem 266: 5891-5897, 1991.
109. Jideama NM, Crawford BH, Hussain AK, Raynor RL. Dephosphory-
lation specificities of protein phosphatase for cardiac troponin I, tro-
ponin T, and sites within troponin T. Int J Biol Sci 2: 1-9, 2006.
110. Jin JP, Chen A, Ogut O, Huang QQ. Conformational modulation of
slow skeletal muscle troponin T by an NH(2)-terminal metal-binding
extension. Am J Physiol Cell Physiol 279: C1067-C1077, 2000.
111. Jin JP, Wang K. Cloning, expression, and protein interaction of human
nebulin fragments composed of varying numbers of sequence modules.
J Biol Chem 266: 21215-21223, 1991.
112. Jin JP, Wang K. Nebulin as a giant actin-binding template protein
in skeletal muscle sarcomere. Interaction of actin and cloned human
nebulin fragments. FEBS Lett 281: 93-96, 1991.
113. Kabsch W, Mannherz HG, Suck D, Pai EF, Holmes KC. Atomic struc-
ture of the actin:DNase I complex. Nature 347: 37-44, 1990.
114. Kalyva A, Parthenakis FI, Marketou ME, Kontaraki JE, Vardas PE.
Biochemical characterisation of Troponin C mutations causing hyper-
trophic and dilated cardiomyopathies. J Muscle Res Cell Motil 35:
161-178, 2014.
115. Kaski JP, Syrris P, Burch M, Tome-Esteban MT, Fenton M, Chris-
tiansen M, Andersen PS, Sebire N, Ashworth M, Deanfield JE,
McKenna WJ, Elliott PM. Idiopathic restrictive cardiomyopathy in
children is caused by mutations in cardiac sarcomere protein genes.
Heart 94: 1478-1484, 2008.
116. Kazmierski ST, Antin PB, Witt CC, Huebner N, McElhinny AS,
Labeit S, Gregorio CC. The complete mouse nebulin gene sequence
and the identification of cardiac nebulin. J Mol Biol 328: 835-846,
2003.
117. Kentish JC, McCloskey DT, Layland J, Palmer S, Leiden JM, Mar-
tin AF, Solaro RJ. Phosphorylation of troponin I by protein kinase A
accelerates relaxation and crossbridge cycle kinetics in mouse ventric-
ular muscle. Circ Res 88: 1059-1065, 2001.
118. Kinbara K, Sorimachi H, Ishiura S, Suzuki K. Muscle-specific calpain,
p94, interacts with the extreme C-terminal region of connectin, a unique
region flanked by two immunoglobulin C2 motifs. Arch Biochem Bio-
phys 342: 99-107, 1997.
119. Kinosian HJ, Selden LA, Estes JE, Gershman LC. Actin filament
annealing in the presence of ATP and phalloidin. Biochemistry 32:
12353-12357, 1993.
120. Kinosian HJ, Selden LA, Estes JE, Gershman LC. Nucleotide binding
to actin. Cation dependence of nucleotide dissociation and exchange
rates. J Biol Chem 268: 8683-8691, 1993.
121. Kleerekoper Q, Howarth JW, Guo X, Solaro RJ, Rosevear PR. Car-
diac troponin I induced conformational changes in cardiac troponin C
as monitored by NMR using site-directed spin and isotope labeling.
Biochemistry 34: 13343-13352, 1995.
122. Knoll R, Buyandelger B, Lab M. The sarcomeric Z-disc and Z-
discopathies. J Biomed Biotechnol 2011: 569628, 2011.
123. Kolh P, Windecker S, Alfonso F, Collet JP, Cremer J, Falk V, Filip-
patos G, Hamm C, Head SJ, Juni P, Kappetein AP, Kastrati A, Knuuti J,
Landmesser U, Laufer G, Neumann FJ, Richter DJ, Schauerte P, Sousa
Uva M, Stefanini GG, Taggart DP, Torracca L, Valgimigli M, Wijns
W, Witkowski A, Zamorano JL, Achenbach S, Baumgartner H, Bax JJ,
Bueno H, Dean V, Deaton C, Erol C, Fagard R, Ferrari R, Hasdai D,
Hoes AW, Kirchhof P, Lancellotti P, Linhart A, Nihoyannopoulos P,
Piepoli MF, Ponikowski P, Sirnes PA, Tamargo JL, Tendera M, Tor-
bicki A, Pepper J, Anyanwu A, Badimon L, Bauersachs J, Baumbach
A, Beygui F, Bonaros N, De Carlo M, Dobrev D, Dunning J, Eeckhout
E, Gielen S, Luckraz H, Mahrholdt H, Montalescot G, Paparella D,
Rastan AJ, Sanmartin M, Sergeant P, Silber S, Tamargo J, ten Berg
J, Thiele H, van Geuns RJ, Wagner HO, Wassmann S, Wendler O.
2014 ESC/EACTS Guidelines on myocardial revascularization: The
Task Force on Myocardial Revascularization of the European Soci-
ety of Cardiology (ESC) and the European Association for Cardio-
Thoracic Surgery (EACTS). Developed with the special contribution
of the European Association of Percutaneous Cardiovascular Interven-
tions (EAPCI). Eur J Cardio-Thorac 46: 517-592, 2014.
124. Kontrogianni-Konstantopoulos A, Catino DH, Strong JC, Sutter S,
Borisov AB, Pumplin DW, Russell MW, Bloch RJ. Obscurin modu-
lates the assembly and organization of sarcomeres and the sarcoplasmic
reticulum. FASEB J 20: 2102-2111, 2006.
125. Kontrogianni-Konstantopoulos A, Jones EM, Van Rossum DB, Bloch
RJ. Obscurin is a ligand for small ankyrin 1 in skeletal muscle. Mol
Biol Cell 14: 1138-1148, 2003.
Volume 7, April 2017
Myofilament Proteins in Myocyte Contraction
126. Koretz JF, Irving TC, Wang K. Filamentous aggregates of native titin
and binding of C-protein and AMP-deaminase. Arch Biochem Biophys
304: 305-309, 1993.
127. Krendel M, Mooseker MS. Myosins: Tails (and heads) of functional
diversity. Physiology (Bethesda) 20: 239-251, 2005.
128. Krenz M, Robbins J. Impact of beta-myosin heavy chain expression
on cardiac function during stress. J Am Coll Cardiol 44: 2390-2397,
2004.
129. Kruger M, Kotter S, Grutzner A, Lang P, Andresen C, Redfield MM,
Butt E, dos Remedios CG, Linke WA. Protein kinase G modulates
human myocardial passive stiffness by phosphorylation of the titin
springs. Circ Res 104: 87-94, 2009.
130. Kruger M, Linke WA. Protein kinase-A phosphorylates titin in human
heart muscle and reduces myofibrillar passive tension. J Muscle Res
Cell Motil 27: 435-444, 2006.
131. Kühne W. Untersuchungen über das Protoplasma und die Contractili-
tat. Leipzig, Engelmann, 1864.
132. Kulikovskaya I, McClellan G, Flavigny J, Carrier L, Winegrad S. Effect
of MyBP-C binding to actin on contractility in heart muscle. J Gen
Physiol 122: 761-774, 2003.
133. Kunst G, Kress KR, Gruen M, Uttenweiler D, Gautel M, Fink RH.
Myosin binding protein C, a phosphorylation-dependent force regulator
in muscle that controls the attachment of myosin heads by its interaction
with myosin S2. Circ Res 86: 51-58, 2000.
134. Kuster DW, Bawazeer AC, Zaremba R, Goebel M, Boontje NM, van
der Velden J. Cardiac myosin binding protein C phosphorylation in
cardiac disease. J Muscle Res Cell Motil 33: 43-52, 2012.
135. Kuster DW, Govindan S, Springer TI, Martin JL, Finley NL, Sadayap-
pan S. A hypertrophic cardiomyopathy-associated MYBPC3 mutation
common in populations of South Asian descent causes contractile dys-
function. J Biol Chem 290: 5855-5867, 2015.
136. Kuster DW, Sequeira V, Najafi A, Boontje NM, Wijnker PJ, Witjas-
Paalberends ER, Marston SB, Dos Remedios CG, Carrier L, Demmers
JA, Redwood C, Sadayappan S, van der Velden J. GSK3beta phosphory-
lates newly identified site in the proline-alanine-rich region of cardiac
myosin-binding protein C and alters cross-bridge cycling kinetics in
human: Short communication. Circ Res 112: 633-639, 2013.
137. Labeit D, Watanabe K, Witt C, Fujita H, Wu Y, Lahmers S, Funck T,
Labeit S, Granzier H. Calcium-dependent molecular spring elements
in the giant protein titin. Proc Natl Acad Sci U S A 100: 13716-13721,
2003.
138. Labeit S, Gautel M, Lakey A, Trinick J. Towards a molecular under-
standing of titin. EMBO J 11: 1711-1716, 1992.
139. Labeit S, Gibson T, Lakey A, Leonard K, Zeviani M, Knight P, Wardale
J, Trinick J. Evidence that nebulin is a protein-ruler in muscle thin
filaments. FEBS Lett 282: 313-316, 1991.
140. Labeit S, Kolmerer B. The complete primary structure of human nebulin
and its correlation to muscle structure. J Mol Biol 248: 308-315, 1995.
141. Labeit S, Kolmerer B. Titins: Giant proteins in charge of muscle ultra-
structure and elasticity. Science 270: 293-296, 1995.
142. Labeit S, Lahmers S, Burkart C, Fong C, McNabb M, Witt S, Witt C,
Labeit D, Granzier H. Expression of distinct classes of titin isoforms in
striated and smooth muscles by alternative splicing, and their conserved
interaction with filamins. J Mol Biol 362: 664-681, 2006.
143. Lahmers S, Wu Y, Call DR, Labeit S, Granzier H. Developmental
control of titin isoform expression and passive stiffness in fetal and
neonatal myocardium. Circ Res 94: 505-513, 2004.
144. Lange S, Auerbach D, McLoughlin P, Perriard E, Schafer BW, Perriard
JC, Ehler E. Subcellular targeting of metabolic enzymes to titin in heart
muscle may be mediated by DRAL/FHL-2. J Cell Sci 115: 4925-4936,
2002.
145. Lange S, Ouyang K, Meyer G, Cui L, Cheng H, Lieber RL, Chen J.
Obscurin determines the architecture of the longitudinal sarcoplasmic
reticulum. J Cell Sci 122: 2640-2650, 2009.
146. Lange S, Perera S, Teh P, Chen J. Obscurin and KCTD6 regulate cullin-
dependent small ankyrin-1 (sAnk1.5) protein turnover. Mol Biol Cell
23: 2490-2504, 2012.
147. Lee EH, Gao M, Pinotsis N, Wilmanns M, Schulten K. Mechanical
strength of the titin Z1Z2-telethonin complex. Structure 14: 497-509,
2006.
148. Lehrer SS, Qian YD, Hvidt S. Assembly of the native heterodimer of
Rana esculenta tropomyosin by chain exchange. Science 246: 926-928,
1989.
149. Lin B, Govindan S, Lee K, Zhao P, Han R, Runte KE, Craig R, Palmer
BM, Sadayappan S. Cardiac Myosin binding protein-C plays no reg-
ulatory role in skeletal muscle structure and function. PLOS One 8:
e69671, 2013.
150. Linke WA. Titin elasticity in the context of the sarcomere: Force and
extensibility measurements on single myofibrils. Adv Exp Med Biol
481: 179-202; discussion 203-176, 2000.
151. Linke WA, Ivemeyer M, Labeit S, Hinssen H, Ruegg JC, Gautel M.
Actin-titin interaction in cardiac myofibrils: Probing a physiological
role. Biophys J 73: 905-919, 1997.
689
Myofilament Proteins in Myocyte Contraction
152. Linke WA, Rudy DE, Centner T, Gautel M, Witt C, Labeit S, Gregorio
CC. I-band titin in cardiac muscle is a three-element molecular spring
and is critical for maintaining thin filament structure. J Cell Biol 146:
631-644, 1999.
153. Lompre AM, Nadal-Ginard B, Mahdavi V. Expression of the cardiac
ventricular alpha- and beta-myosin heavy chain genes is developmen-
tally and hormonally regulated. J Biol Chem 259: 6437-6446, 1984.
154. Lorenz M, Poole KJ, Popp D, Rosenbaum G, Holmes KC. An atomic
model of the unregulated thin filament obtained by X-ray fiber diffrac-
tion on oriented actin-tropomyosin gels. J Mol Biol 246: 108-119, 1995.
155. Luther PK, Craig R. Modulation of striated muscle contraction by
binding of myosin binding protein C to actin. Bioarchitecture 1: 277-
283, 2011.
156. Luther PK, Winkler H, Taylor K, Zoghbi ME, Craig R, Padron R, Squire
JM, Liu J. Direct visualization of myosin-binding protein C bridging
myosin and actin filaments in intact muscle. Proc Natl Acad Sci U S A
108: 11423-11428, 2011.
157. Mahdavi V, Chambers AP, Nadal-Ginard B. Cardiac alpha- and beta-
myosin heavy chain genes are organized in tandem. Proc Natl Acad Sci
U S A 81: 2626-2630, 1984.
158. Mahdavi V, Lompre AM, Chambers AP, Nadal-Ginard B. Cardiac
myosin heavy chain isozymic transitions during development and under
pathological conditions are regulated at the level of mRNA availability.
Eur Heart J 5(Suppl F): 181-191, 1984.
159. Makarenko I, Opitz CA, Leake MC, Neagoe C, Kulke M, Gwathmey
JK, del Monte F, Hajjar RJ, Linke WA. Passive stiffness changes caused
by upregulation of compliant titin isoforms in human dilated cardiomy-
opathy hearts. Circ Res 95: 708-716, 2004.
160. Marston S, Montgiraud C, Munster AB, Copeland O, Choi O, Dos
Remedios C, Messer AE, Ehler E, Knoll R. OBSCN mutations associ-
ated with dilated cardiomyopathy and haploinsufficiency. PLOS ONE
10: e0138568, 2015.
161. Maruyama K. Connectin, an elastic protein from myofibrils. J Biochem
80: 405-407, 1976.
162. Maruyama K, Kimura S, Kuroda M, Handa S. Connectin, an elastic
protein of muscle. Its abundance in cardiac myofibrils. J Biochem 82:
347-350, 1977.
163. Maruyama K, Matsubara S, Natori R, Nonomura Y, Kimura S. Con-
nectin, an elastic protein of muscle. Characterization and function.
J Biochem 82: 317-337, 1977.
164. Mastrototaro G, Liang X, Li X, Carullo P, Piroddi N, Tesi C, Gu Y,
Dalton ND, Peterson KL, Poggesi C, Sheikh F, Chen J, Bang ML.
Nebulette knockout mice have normal cardiac function, but show Z-
line widening and up-regulation of cardiac stress markers. Cardiovasc
Res 107: 216-225, 2015.
165. Mateja RD, Greaser ML, de Tombe PP. Impact of titin isoform on
length dependent activation and cross-bridge cycling kinetics in rat
skeletal muscle. Biochim Biophys Acta 1833: 804-811, 2013.
166. Matsumoto Y, Hayashi T, Inagaki N, Takahashi M, Hiroi S, Naka-
mura T, Arimura T, Nakamura K, Ashizawa N, Yasunami M, Ohe T,
Yano K, Kimura A. Functional analysis of titin/connectin N2-B muta-
tions found in cardiomyopathy. J Muslce Res Cell Motil 26: 367-374,
2005.
167. Mayans O, van der Ven PF, Wilm M, Mues A, Young P, Furst DO,
Wilmanns M, Gautel M. Structural basis for activation of the titin
kinase domain during myofibrillogenesis. Nature 395: 863-869, 1998.
168. McConnell BK, Jones KA, Fatkin D, Arroyo LH, Lee RT, Aristizabal O,
Turnbull DH, Georgakopoulos D, Kass D, Bond M, Niimura H, Schoen
FJ, Conner D, Fischman DA, Seidman CE, Seidman JG. Dilated car-
diomyopathy in homozygous myosin-binding protein-C mutant mice.
J Clin Invest 104: 1771, 1999.
169. McElhinny AS, Schwach C, Valichnac M, Mount-Patrick S, Gregorio
CC. Nebulin regulates the assembly and lengths of the thin filaments in
striated muscle. J Cell Biol 170: 947-957, 2005.
170. McKillop DF, Geeves MA. Regulation of the interaction between actin
and myosin subfragment 1: Evidence for three states of the thin filament.
Biophys J 65: 693-701, 1993.
171. McLachlan AD, Stewart M. Tropomyosin coiled-coil interactions: Evi-
dence for an unstaggered structure. J Mol Biol 98: 293-304, 1975.
172. McLaughlin PJ, Gooch JT, Mannherz HG, Weeds AG. Structure of gel-
solin segment 1-actin complex and the mechanism of filament severing.
Nature 364: 685-692, 1993.
173. Michele DE, Metzger JM. Physiological consequences of tropomyosin
mutations associated with cardiac and skeletal myopathies. J Mol Med
(Berl) 78: 543-553, 2000.
174. Millevoi S, Trombitas K, Kolmerer B, Kostin S, Schaper J, Pelin K,
Granzier H, Labeit S. Characterization of nebulette and nebulin and
emerging concepts of their roles for vertebrate Z-discs. J Mol Biol 282:
111-123, 1998.
175. Milligan RA. Protein-protein interactions in the rigor actomyosin com-
plex. Proc Natl Acad Sci U S A 93: 21-26, 1996.
176. Milligan RA, Whittaker M, Safer D. Molecular structure of F-actin and
location of surface binding sites. Nature 348: 217-221, 1990.
690
Comprehensive Physiology
177. Miyamoto CA, Fischman DA, Reinach FC. The interface between
MyBP-C and myosin: Site-directed mutagenesis of the CX myosin-
binding domain of MyBP-C. J Muscle Res Cell Motil 20: 703-715,
1999.
178. Miyata S, Minobe W, Bristow MR, Leinwand LA. Myosin heavy chain
isoform expression in the failing and nonfailing human heart. Circ Res
86: 386-390, 2000.
179. Mogensen J, Hey T, Lambrecht S. A systematic review of phenotypic
features associated with cardiac troponin I mutations in hereditary car-
diomyopathies. Can J Cardiol 31: 1377-1385, 2015.
180. Mohamed AS, Dignam JD, Schlender KK. Cardiac myosin-binding
protein C (MyBP-C): Identification of protein kinase A and protein
kinase C phosphorylation sites. Arch Biochem Biophys 358: 313-319,
1998.
181. Molkentin JD, Jobe SM, Markham BE. Alpha-myosin heavy chain
gene regulation: Delineation and characterization of the cardiac muscle-
specific enhancer and muscle-specific promoter. J Mol Cell Cardiol 28:
1211-1225, 1996.
182. Moncman CL, Wang K. Nebulette: A 107 kD nebulin-like protein in
cardiac muscle. Cell Motil Cytoskeleton 32: 205-225, 1995.
183. Moncman CL, Wang K. Targeted disruption of nebulette protein expres-
sion alters cardiac myofibril assembly and function. Exp Cell Res 273:
204-218, 2002.
184. Moore JR, Leinwand L, Warshaw DM. Understanding cardiomyopathy
phenotypes based on the functional impact of mutations in the myosin
motor. Circ Res 111: 375-385, 2012.
185. Moos C, Offer G, Starr R, Bennett P. Interaction of C-protein with
myosin, myosin rod and light meromyosin. J Mol Biol 97: 1-9,
1975.
186. Morgan JE, Partridge TA. Muscle satellite cells. Int J Biochem Cell Biol
35: 1151-1156, 2003.
187. Morris EP, Lehrer SS. Troponin-tropomyosin interactions. Fluores-
cence studies of the binding of troponin, troponin T, and chymotryptic
troponin T fragments to specifically labeled tropomyosin. Biochemistry
23: 2214-2220, 1984.
188. Mues A, van der Ven PF, Young P, Furst DO, Gautel M. Two
immunoglobulin-like domains of the Z-disc portion of titin interact in a
conformation-dependent way with telethonin. FEBS Lett 428: 111-114,
1998.
189. Mun JY, Previs MJ, Yu HY, Gulick J, Tobacman LS, Beck Previs S,
Robbins J, Warshaw DM, Craig R. Myosin-binding protein C displaces
tropomyosin to activate cardiac thin filaments and governs their speed
by an independent mechanism. Proc Natl Acad Sci U S A 111: 2170-
2175, 2014.
190. Muthuchamy M, Grupp IL, Grupp G, O’Toole BA, Kier AB, Boivin
GP, Neumann J, Wieczorek DF. Molecular and physiological effects
of overexpressing striated muscle beta-tropomyosin in the adult murine
heart. J Biol Chem 270: 30593-30603, 1995.
191. Nadal-Ginard B, Mahdavi V. Molecular basis of cardiac perfor-
mance. Plasticity of the myocardium generated through protein isoform
switches. J Clin Invest 84: 1693-1700, 1989.
192. Nagueh SF, Shah G, Wu Y, Torre-Amione G, King NM, Lahmers
S, Witt CC, Becker K, Labeit S, Granzier HL. Altered titin expres-
sion, myocardial stiffness, and left ventricular function in patients with
dilated cardiomyopathy. Circulation 110: 155-162, 2004.
193. Neagoe C, Kulke M, del Monte F, Gwathmey JK, de Tombe PP, Hajjar
RJ, Linke WA. Titin isoform switch in ischemic human heart disease.
Circulation 106: 1333-1341, 2002.
194. Noland TA, Jr., Kuo JF. Protein kinase C phosphorylation of cardiac
troponin I or troponin T inhibits Ca2(+)-stimulated actomyosin MgAT-
Pase activity. J Biol Chem 266: 4974-4978, 1991.
195. Noland TA, Jr., Kuo JF. Protein kinase C phosphorylation of cardiac
troponin T decreases Ca(2+)-dependent actomyosin MgATPase activ-
ity and troponin T binding to tropomyosin-F-actin complex. Biochem
J 288(Pt 1): 123-129, 1992.
196. Nyland LR, Palmer BM, Chen Z, Maughan DW, Seidman CE, Seidman
JG, Kreplak L, Vigoreaux JO. Cardiac myosin binding protein-C is
essential for thick-filament stability and flexural rigidity. Biochem J
96: 3273-3280, 2009.
197. O’Brien PJ, Smith DE, Knechtel TJ, Marchak MA, Pruimboom-Brees
I, Brees DJ, Spratt DP, Archer FJ, Butler P, Potter AN, Provost JP,
Richard J, Snyder PA, Reagan WJ. Cardiac troponin I is a sensitive,
specific biomarker of cardiac injury in laboratory animals. Lab Anim
40: 153-171, 2006.
198. Obermann WM, Gautel M, Weber K, Furst DO. Molecular structure of
the sarcomeric M band: Mapping of titin and myosin binding domains
in myomesin and the identification of a potential regulatory phospho-
rylation site in myomesin. EMBO J 16: 211-220, 1997.
199. Offer G, Moos C, Starr R. A new protein of the thick filaments of verte-
brate skeletal myofibrils. Extractions, purification and characterization.
J Mol Biol 74: 653-676, 1973.
200. Ohtsuki I, Maruyama K, Ebashi S. Regulatory and cytoskeletal proteins
of vertebrate skeletal muscle. Adv Protein Chem 38: 1-67, 1986.
Volume 7, April 2017
Comprehensive Physiology
201. Okagaki T, Weber FE, Fischman DA, Vaughan KT, Mikawa T, Reinach
FC. The major myosin-binding domain of skeletal muscle MyBP-C
(C protein) resides in the COOH-terminal, immunoglobulin C2 motif.
J Cell Biol 123: 619-626, 1993.
202. Okazaki K, Holtzer H. Myogenesis: Fusion, myosin synthesis, and the
mitotic cycle. Proc Natl Acad Sci U S A 56: 1484-1490, 1966.
203. Opitz CA, Leake MC, Makarenko I, Benes V, Linke WA. Developmen-
tally regulated switching of titin size alters myofibrillar stiffness in the
perinatal heart. Circ Res 94: 967-975, 2004.
204. Orlova A, Egelman EH. Structural dynamics of F-actin: I. Changes in
the C terminus. J Mol Biol 245: 582-597, 1995.
205. Orlova A, Prochniewicz E, Egelman EH. Structural dynamics of F-
actin: II. Cooperativity in structural transitions. J Mol Biol 245: 598-
607, 1995.
206. Pacchiarotti I, Bond DJ, Baldessarini RJ, Nolen WA, Grunze H, Licht
RW, Post RM, Berk M, Goodwin GM, Sachs GS, Tondo L, Findling
RL, Youngstrom EA, Tohen M, Undurraga J, Gonzalez-Pinto A, Gold-
berg JF, Yildiz A, Altshuler LL, Calabrese JR, Mitchell PB, Thase
ME, Koukopoulos A, Colom F, Frye MA, Malhi GS, Fountoulakis KN,
Vazquez G, Perlis RH, Ketter TA, Cassidy F, Akiskal H, Azorin JM,
Valenti M, Mazzei DH, Lafer B, Kato T, Mazzarini L, Martinez-Aran
A, Parker G, Souery D, Ozerdem A, McElroy SL, Girardi P, Bauer
M, Yatham LN, Zarate CA, Nierenberg AA, Birmaher B, Kanba S,
El-Mallakh RS, Serretti A, Rihmer Z, Young AH, Kotzalidis GD, Mac-
Queen GM, Bowden CL, Ghaemi SN, Lopez-Jaramillo C, Rybakowski
J, Ha K, Perugi G, Kasper S, Amsterdam JD, Hirschfeld RM, Kapczin-
ski F, Vieta E. The International Society for Bipolar Disorders (ISBD)
task force report on antidepressant use in bipolar disorders. Am J Psy-
chiatry 170: 1249-1262, 2013.
207. Pappas CT, Bhattacharya N, Cooper JA, Gregorio CC. Nebulin interacts
with CapZ and regulates thin filament architecture within the Z-disc.
Mol Biol Cell 19: 1837-1847, 2008.
208. Pappas CT, Bliss KT, Zieseniss A, Gregorio CC. The Nebulin family:
An actin support group. Trends Cell Biol 21: 29-37, 2011.
209. Pappas CT, Krieg PA, Gregorio CC. Nebulin regulates actin fila-
ment lengths by a stabilization mechanism. J Cell Biol 189: 859-870,
2010.
210. Pearlstone JR, Smillie LB. Binding of troponin-T fragments to several
types of tropomyosin. Sensitivity to Ca2+ in the presence of troponin-
C. J Biol Chem 257: 10587-10592, 1982.
211. Peng J, Raddatz K, Labeit S, Granzier H, Gotthardt M. Muscle atrophy
in titin M-line deficient mice. J Muscle Res Cell Motil 26: 381-388,
2005.
212. Perry NA, Vitolo MI, Martin SS, Kontrogianni-Konstantopoulos A.
Loss of the obscurin-RhoGEF downregulates RhoA signaling and
increases microtentacle formation and attachment of breast epithelial
cells. Oncotarget 5: 8558-8568, 2014.
213. Perry SV. Vertebrate tropomyosin: Distribution, properties and func-
tion. J Muscle Res Cell Motil 22: 5-49, 2001.
214. Pfuhl M, Gautel M. Structure, interactions and function of the N-
terminus of cardiac myosin binding protein C (MyBP-C): Who does
what, with what, and to whom? J Muscle Res Cell Motil 33: 83-94,
2012.
215. Pope B, Hoh JF, Weeds A. The ATPase activities of rat cardiac myosin
isoenzymes. FEBS Lett 118: 205-208, 1980.
216. Potter JD, Gergely J. Troponin, tropomyosin, and actin interactions
in the Ca2+ regulation of muscle contraction. Biochemistry 13: 2697-
2703, 1974.
217. Potter JD, Sheng Z, Pan BS, Zhao J. A direct regulatory role for troponin
T and a dual role for troponin C in the Ca2+ regulation of muscle
contraction. J Biol Chem 270: 2557-2562, 1995.
218. Previs MJ, Beck Previs S, Gulick J, Robbins J, Warshaw DM. Molec-
ular mechanics of cardiac myosin-binding protein C in native thick
filaments. Science 337: 1215-1218, 2012.
219. Previs MJ, Mun JY, Michalek AJ, Previs SB, Gulick J, Robbins J,
Warshaw DM, Craig R. Phosphorylation and calcium antagonistically
tune myosin-binding protein C’s structure and function. Proc Natl Acad
Sci U S A 113: 3239-3244, 2016.
220. Puchner EM, Alexandrovich A, Kho AL, Hensen U, Schafer LV, Brand-
meier B, Grater F, Grubmuller H, Gaub HE, Gautel M. Mechanoen-
zymatics of titin kinase. Proc Natl Acad Sci U S A 105: 13385-13390,
2008.
221. Pyle WG, Solaro RJ. At the crossroads of myocardial signaling: The
role of Z-discs in intracellular signaling and cardiac function. Circ Res
94: 296-305, 2004.
222. Quiat D, Voelker KA, Pei J, Grishin NV, Grange RW, Bassel-Duby R,
Olson EN. Concerted regulation of myofiber-specific gene expression
and muscle performance by the transcriptional repressor Sox6. Proc
Natl Acad Sci U S A 108: 10196-10201, 2011.
223. Raeker MO, Bieniek AN, Ryan AS, Tsai HJ, Zahn KM, Russell MW.
Targeted deletion of the zebrafish obscurin A RhoGEF domain affects
heart, skeletal muscle and brain development. Dev Biol 337: 432-443,
2010.
Volume 7, April 2017
Myofilament Proteins in Myocyte Contraction
224. Raeker MO, Su F, Geisler SB, Borisov AB, Kontrogianni-
Konstantopoulos A, Lyons SE, Russell MW. Obscurin is required for
the lateral alignment of striated myofibrils in zebrafish. Dev Dyn 235:
2018-2029, 2006.
225. Randazzo D, Giacomello E, Lorenzini S, Rossi D, Pierantozzi E,
Blaauw B, Reggiani C, Lange S, Peter AK, Chen J, Sorrentino V.
Obscurin is required for ankyrinB-dependent dystrophin localization
and sarcolemma integrity. J Cell Biol 200: 523-536, 2013.
226. Raskin A, Lange S, Banares K, Lyon RC, Zieseniss A, Lee LK,
Yamazaki KG, Granzier HL, Gregorio CC, McCulloch AD, Omens
JH, Sheikh F. A novel mechanism involving four-and-a-half LIM
domain protein-1 and extracellular signal-regulated kinase-2 regulates
titin phosphorylation and mechanics. J Biol Chem 287: 29273-29284,
2012.
227. Ratti J, Rostkova E, Gautel M, Pfuhl M. Structure and interactions of
myosin-binding protein C domain C0: Cardiac-specific regulation of
myosin at its neck? J Biol Chem 286: 12650-12658, 2011.
228. Rayment I, Holden HM, Whittaker M, Yohn CB, Lorenz M, Holmes
KC, Milligan RA. Structure of the actin-myosin complex and its impli-
cations for muscle contraction. Science 261: 58-65, 1993.
229. Rayment I, Rypniewski WR, Schmidt-Base K, Smith R, Tomchick DR,
Benning MM, Winkelmann DA, Wesenberg G, Holden HM. Three-
dimensional structure of myosin subfragment-1: A molecular motor.
Science 261: 50-58, 1993.
230. Razumova MV, Bezold KL, Tu AY, Regnier M, Harris SP. Contri-
bution of the myosin binding protein C motif to functional effects in
permeabilized rat trabeculae. J Gen Physiol 132: 575-585, 2008.
231. Razumova MV, Shaffer JF, Tu AY, Flint GV, Regnier M, Harris SP.
Effects of the N-terminal domains of myosin binding protein-C in an
in vitro motility assay: Evidence for long-lived cross-bridges. J Biol
Chem 281: 35846-35854, 2006.
232. Robertson SP, Johnson JD, Holroyde MJ, Kranias EG, Potter JD, Solaro
RJ. The effect of troponin I phosphorylation on the Ca2+-binding
properties of the Ca2+-regulatory site of bovine cardiac troponin. J Biol
Chem 257: 260-263, 1982.
233. Robinson BF, Epstein SE, Beiser GD, Braunwald E. Control of heart
rate by the autonomic nervous system. Studies in man on the inter-
relation between baroreceptor mechanisms and exercise. Circ Res 19:
400-411, 1966.
234. Robinson P, Griffiths PJ, Watkins H, Redwood CS. Dilated and hyper-
trophic cardiomyopathy mutations in troponin and alpha-tropomyosin
have opposing effects on the calcium affinity of cardiac thin filaments.
Circ Res 101: 1266-1273, 2007.
235. Rome E, Offer G, Pepe FA. X-ray diffraction of muscle labelled with
antibody to C-protein. Nat New Biol 244: 152-154, 1973.
236. Rosol M, Lehman W, Craig R, Landis C, Butters C, Tobacman LS.
Three-dimensional reconstruction of thin filaments containing mutant
tropomyosin. Biophys J 78: 908-917, 2000.
237. Russell MW, Raeker MO, Korytkowski KA, Sonneman KJ. Identi-
fication, tissue expression and chromosomal localization of human
Obscurin-MLCK, a member of the titin and Dbl families of myosin
light chain kinases. Gene 282: 237-246, 2002.
238. Sadayappan S, de Tombe PP. Cardiac myosin binding protein-C:
Redefining its structure and function. Biophys Rev 4: 93-106, 2012.
239. Satoh M, Takahashi M, Sakamoto T, Hiroe M, Marumo F, Kimura A.
Structural analysis of the titin gene in hypertrophic cardiomyopathy:
Identification of a novel disease gene. Biochem Biophys Res Commun
262: 411-417, 1999.
240. Satyshur KA, Rao ST, Pyzalska D, Drendel W, Greaser M, Sundar-
alingam M. Refined structure of chicken skeletal muscle troponin C in
the two-calcium state at 2-A resolution. J Biol Chem 263: 1628-1647,
1988.
241. Schaertl S, Lehrer SS, Geeves MA. Separation and characterization of
the two functional regions of troponin involved in muscle thin filament
regulation. Biochemistry 34: 15890-15894, 1995.
242. Schiaffino S, Reggiani C. Fiber types in mammalian skeletal muscles.
Physiol Rev 91: 1447-1531, 2011.
243. Schutt CE, Lindberg U, Myslik J, Strauss N. Molecular packing in
profilin: Actin crystals and its implications. J Mol Biol 209: 735-746,
1989.
244. Severs NJ. The cardiac muscle cell. Bioessays 22: 188-199, 2000.
245. Shaffer JF, Harris SP. Species-specific differences in the Pro-Ala rich
region of cardiac myosin binding protein-C. J Muscle Res Cell Motil
30: 303-306, 2009.
246. Sia SK, Li MX, Spyracopoulos L, Gagne SM, Liu W, Putkey JA, Sykes
BD. Structure of cardiac muscle troponin C unexpectedly reveals a
closed regulatory domain. J Biol Chem 272: 18216-18221, 1997.
247. Spudich JA. The myosin mesa and a possible unifying hypothesis for
the molecular basis of human hypertrophic cardiomyopathy. Biochem
Soc Trans 43: 64-72, 2015.
248. Squire JM, Luther PK, Knupp C. Structural evidence for the interaction
of C-protein (MyBP-C) with actin and sequence identification of a
possible actin-binding domain. J Mol Biol 331: 713-724, 2003.
691
Myofilament Proteins in Myocyte Contraction
249. Starr R, Offer G. The interaction of C-protein with heavy meromyosin
and subfragment-2. Biochem J 171: 813-816, 1978.
250. Straub FB. Actin, II. Stud Inst Med Chem Univ Szeged III: 23-37,
1943.
251. Sumandea MP, Pyle WG, Kobayashi T, de Tombe PP, Solaro RJ.
Identification of a functionally critical protein kinase C phosphorylation
residue of cardiac troponin T. J Biol Chem 278: 35135-35144, 2003.
252. Sutter SB, Raeker MO, Borisov AB, Russell MW. Orthologous rela-
tionship of obscurin and Unc-89: Phylogeny of a novel family of tandem
myosin light chain kinases. Dev Genes Evol 214: 352-359, 2004.
253. Suzuki T, Palmer BM, James J, Wang Y, Chen Z, VanBuren P, Maughan
DW, Robbins J, LeWinter MM. Effects of cardiac myosin isoform vari-
ation on myofilament function and crossbridge kinetics in transgenic
rabbits. Circ Heart Fail 2: 334-341, 2009.
254. Syska H, Wilkinson JM, Grand RJ, Perry SV. The relationship between
biological activity and primary structure of troponin I from white skele-
tal muscle of the rabbit. Biochem J 153: 375-387, 1976.
255. Tajsharghi H, Ohlsson M, Palm L, Oldfors A. Myopathies associated
with beta-tropomyosin mutations. Neuromuscul Disord 22: 923-933,
2012.
256. Takeda S, Yamashita A, Maeda K, Maeda Y. Structure of the core
domain of human cardiac troponin in the Ca(2+)-saturated form. Nature
424: 35-41, 2003.
257. Tanigawa G, Jarcho JA, Kass S, Solomon SD, Vosberg HP, Seidman
JG, Seidman CE. A molecular basis for familial hypertrophic cardiomy-
opathy: An alpha/beta cardiac myosin heavy chain hybrid gene. Cell
62: 991-998, 1990.
258. Tardiff JC, Hewett TE, Factor SM, Vikstrom KL, Robbins J, Leinwand
LA. Expression of the beta (slow)-isoform of MHC in the adult mouse
heart causes dominant-negative functional effects. Am J Physiol Heart
Circ Physiol 278: H412-H419, 2000.
259. Terui T, Sodnomtseren M, Matsuba D, Udaka J, Ishiwata S, Ohtsuki I,
Kurihara S, Fukuda N. Troponin and titin coordinately regulate length-
dependent activation in skinned porcine ventricular muscle. J Gen Phys-
iol 131: 275-283, 2008.
260. Tian LF, Li HY, Jin BF, Pan X, Man JH, Zhang PJ, Li WH, Liang B,
Liu H, Zhao J, Gong WL, Zhou T, Zhang XM. MDM2 interacts with
and downregulates a sarcomeric protein, TCAP. Biochem Bioph Res
Co 345: 355-361, 2006.
261. Tonino P, Pappas CT, Hudson BD, Labeit S, Gregorio CC, Granzier
H. Reduced myofibrillar connectivity and increased Z-disk width in
nebulin-deficient skeletal muscle. J Cell Sci 123: 384-391, 2010.
262. Toyoshima YY, Kron SJ, McNally EM, Niebling KR, Toyoshima C,
Spudich JA. Myosin subfragment-1 is sufficient to move actin filaments
in vitro. Nature 328: 536-539, 1987.
263. Trinick J. Understanding the functions of titin and nebulin. FEBS Lett
307: 44-48, 1992.
264. Trinick J. Titin and nebulin: Protein rulers in muscle? Trends Biochem
Sci 19: 405-409, 1994.
265. Trombitas K, Freiburg A, Centner T, Labeit S, Granzier H. Molecular
dissection of N2B cardiac titin’s extensibility. Biophys J 77: 3189-3196,
1999.
266. Trombitas K, Jin JP, Granzier H. The mechanically active domain of
titin in cardiac muscle. Circ Res 77: 856-861, 1995.
267. van der Ven PF, Furst DO. Assembly of titin, myomesin and M-protein
into the sarcomeric M band in differentiating human skeletal muscle
cells in vitro. Cell Struct Funct 22: 163-171, 1997.
268. Vandekerckhove J, Weber K. The complete amino acid sequence of
actins from bovine aorta, bovine heart, bovine fast skeletal muscle,
and rabbit slow skeletal muscle. A protein-chemical analysis of muscle
actin differentiation. Differentiation 14: 123-133, 1979.
269. Vassylyev DG, Takeda S, Wakatsuki S, Maeda K, Maeda Y. Crystal
structure of troponin C in complex with troponin I fragment at 2.3-A
resolution. Proc Natl Acad Sci U S A 95: 4847-4852, 1998.
270. von der Ecken J, Muller M, Lehman W, Manstein DJ, Penczek PA,
Raunser S. Structure of the F-actin-tropomyosin complex. Nature 519:
114-117, 2015.
271. Vydyanath A, Gurnett CA, Marston S, Luther PK. Axial distribution of
myosin binding protein-C is unaffected by mutations in human cardiac
and skeletal muscle. J Muscle Res Cell Motil 33: 61-74, 2012.
692
Comprehensive Physiology
272. Wang K. Purification of titin and nebulin. Methods Enzymol 85(Pt B):
264-274, 1982.
273. Wang K. Titin/connectin and nebulin: Giant protein rulers of muscle
structure and function. Adv Biophys 33: 123-134, 1996.
274. Wang K, Forbes JG, Jin AJ. Single molecule measurements of titin
elasticity. Prog Biophys Mol Biol 77: 1-44, 2001.
275. Wang K, Knipfer M, Huang QQ, van Heerden A, Hsu LC, Gutierrez
G, Quian XL, Stedman H. Human skeletal muscle nebulin sequence
encodes a blueprint for thin filament architecture. Sequence motifs and
affinity profiles of tandem repeats and terminal SH3. J Biol Chem 271:
4304-4314, 1996.
276. Wang K, McClure J, Tu A. Titin: Major myofibrillar components of
striated muscle. Proc Natl Acad Sci U S A 76: 3698-3702, 1979.
277. Warren CM, Jordan MC, Roos KP, Krzesinski PR, Greaser ML. Titin
isoform expression in normal and hypertensive myocardium. Cardio-
vasc Res 59: 86-94, 2003.
278. Weber FE, Vaughan KT, Reinach FC, Fischman DA. Complete
sequence of human fast-type and slow-type muscle myosin-binding-
protein C (MyBP-C). Differential expression, conserved domain struc-
ture and chromosome assignment. Eur J Biochem 216: 661-669, 1993.
279. White SP, Cohen C, Phillips GN, Jr. Structure of co-crystals of
tropomyosin and troponin. Nature 325: 826-828, 1987.
280. Whitten AE, Jeffries CM, Harris SP, Trewhella J. Cardiac myosin-
binding protein C decorates F-actin: Implications for cardiac function.
Proc Natl Acad Sci U S A 105: 18360-18365, 2008.
281. Witayavanitkul N, Ait Mou Y, Kuster DW, Khairallah RJ, Sarkey J,
Govindan S, Chen X, Ge Y, Rajan S, Wieczorek DF, Irving T, Westfall
MV, de Tombe PP, Sadayappan S. Myocardial infarction-induced N-
terminal fragment of cardiac myosin-binding protein C (cMyBP-C)
impairs myofilament function in human myocardium. J Biol Chem
289: 8818-8827, 2014.
282. Witt CC, Burkart C, Labeit D, McNabb M, Wu Y, Granzier H, Labeit S.
Nebulin regulates thin filament length, contractility, and Z-disk struc-
ture in vivo. EMBO J 25: 3843-3855, 2006.
283. Witt CC, Olivieri N, Centner T, Kolmerer B, Millevoi S, Morell J, Labeit
D, Labeit S, Jockusch H, Pastore A. A survey of the primary structure
and the interspecies conservation of I-band titin’s elastic elements in
vertebrates. J Struct Biol 122: 206-215, 1998.
284. Witt SH, Granzier H, Witt CC, Labeit S. MURF-1 and MURF-2 target
a specific subset of myofibrillar proteins redundantly: Towards under-
standing MURF-dependent muscle ubiquitination. J Mol Biol 350: 713-
722, 2005.
285. Woods EF. The dissociation of tropomyosin by urea. J Mol Biol 16:
581-584, 1966.
286. Woods EF. Molecular weight and subunit structure of tropomyosin B.
J Biol Chem 242: 2859-2871, 1967.
287. Woolner S, Bement WM. Unconventional myosins acting unconven-
tionally. Trends Cell Biol 19: 245-252, 2009.
288. Wu Y, Bell SP, Trombitas K, Witt CC, Labeit S, LeWinter MM,
Granzier H. Changes in titin isoform expression in pacing-induced car-
diac failure give rise to increased passive muscle stiffness. Circulation
106: 1384-1389, 2002.
289. Yamasaki R, Wu Y, McNabb M, Greaser M, Labeit S, Granzier H.
Protein kinase A phosphorylates titin’s cardiac-specific N2B domain
and reduces passive tension in rat cardiac myocytes. Circ Res 90: 1181-
1188, 2002.
290. Yin H, Price F, Rudnicki MA. Satellite cells and the muscle stem cell
niche. Physiol Rev 93: 23-67, 2013.
291. Young P, Ehler E, Gautel M. Obscurin, a giant sarcomeric Rho guanine
nucleotide exchange factor protein involved in sarcomere assembly.
J Cell Biol 154: 123-136, 2001.
292. Young P, Ferguson C, Banuelos S, Gautel M. Molecular structure of
the sarcomeric Z-disk: Two types of titin interactions lead to an asym-
metrical sorting of alpha-actinin. EMBO J 17: 1614-1624, 1998.
293. Zhu C, Yin Z, Ren J, McCormick RJ, Ford SP, Guo W. RBM20 is an
essential factor for thyroid hormone-regulated titin isoform transition.
J Mol Cell Biol 7: 88-90, 2015.
294. Zot AS, Potter JD. Structural aspects of troponin-tropomyosin regula-
tion of skeletal muscle contraction. Annu Rev Biophys Biophys Chem
16: 535-559, 1987.
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