MOLECULAR MOTORS

Ms. M. Banda (B.Pharm, MSc Biochem)

INTRODUCTION
 Movements of cells, organelles and macromolecules are fueled by
energy which is usually derived from ATP.

 Large aggregates of motor proteins undergo cyclic conformational

changes that accumulate into a unified, directional force.

 The interactions among motor proteins, include ionic, hydrogen-

bonding, hydrophobic, and van der Waals interactions at protein
binding sites.

 Motor proteins bring about;

- the contraction of muscles
- the migration of organelles along microtubules,
- the rotation of bacterial flagella and cilia, and
- the movement of some proteins along DNA.

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CONTRACTILE PROTEINS
 Include actin and myosin which make up over 80% of the
protein of muscle mass

 The contractile force of muscle is generated by the interaction

of the two proteins.

 These proteins are arranged in filaments that undergo

transient interactions and slide past each other to bring about
contraction.

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 MYOSIN
 Myosin has six subunits: two heavy chains and four light chains with the heavy
chains accounting for most of the overall structure.

 At their carboxyl termini, the heavy chains are arranged as extended α-helices,
wrapped around each other in a fibrous, left-handed coil whilst at the amino
terminal end, each heavy chain has a globular domain containing an ATP
hydrolyzing site.

 The light chains are associated with the globular domains.

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 When myosin is treated briefly with the protease trypsin, much of the fibrous
tail is cleaved off, dividing the protein into components called light and heavy
meromyosin.

 The globular domain , called myosin subfragment 1, or S1, or simply the myosin
head group makes muscle contraction possible.

 Some exist as one headed and some are two headed.

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Head (motor) domain
 Responsible for generating force.
 Most conserved region in myosin.
 Binds actin and hydrolyses ATP
 ATPase activity is actin – activated.

Tail (effector) domain

 Tail is unique for the myosin type
 Contains binding sites for cell membrane or other tails

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 In muscle cells, molecules of myosin aggregate to form structures called thick
filaments.

 These rod-like structures serve as the core of the contractile unit.

 Within a thick filament, several hundred myosin molecules are arranged with
their fibrous “tails” associated to form a long bipolar structure.

 The globular domains project from either end of this structure, in regular
stacked arrays.

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 ACTIN
 Abundant in almost all eukaryotic cells.

 In muscle, molecules of monomeric actin, called G-actin (globular actin),

associate in a 2 by 2 way to give the appearance of 2 filaments spiraling about
one another in a right handed fashion forming a long polymer called F-actin
(filamentous actin).

 The thin filament consists of F-actin, along with the accessory proteins
troponin and tropomyosin.

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 On addition, of G actin to F actin, each monomer binds ATP, then
hydrolyzes it to ADP.

 This ATP hydrolysis by actin functions only in the assembly of the

filaments; and not for muscle contraction.

 Each actin monomer in the thin filament can bind tightly and
specifically to one myosin head group.

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 Tropomyosin is a rod-shaped fibrous protein and these rods link end to end to
form two helical strands which are wrapped around the F-actin in a longitudinal
fashion.

 Tropomyosin functions to switch the contractile mechanism on and off.

 Troponin is a globular protein made up of three subunits;

 Troponin-T binds troponin to tropomyosin,

 Troponin-C is sensitive to and can reversibly bind to Ca 2+ ions,

 Troponin-I under certain conditions is able to inhibit any interaction between actin
and myosin.

 Both tropomyosin and troponin inhibit actin-myosin interaction in the

absence of Ca2+ ions.

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Molecular structure of muscle
 Skeletal muscle consists of parallel bundles of muscle fibers.

 Muscle fibers are also known as muscle cells or myocytes.

 Each muscle fiber is muliti-nucleated as it is long and can span the

length of the entire muscle

 Each fiber contains about 1,000 myofibrils, each consisting of a vast

number of regularly arrayed thick and thin filaments forming complex
with other proteins.

 A system of flat membranous vesicles called the sarcoplasmic

reticulum surrounds each myofibril.

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 Examined under the electron microscope, muscle fibers reveal alternating
regions of high and low electron density, called the A bands and I bands.

 The A and I bands arise from the arrangement of thick and thin filaments
which are aligned and partially overlapping.

 The I band is the region of the bundle that in cross section would contain
only thin filaments.

 The darker A band stretches the length of the thick filament and includes
the region where parallel thick and thin filaments overlap.

 Bisecting the I band is a thin structure called the Z disk, perpendicular to

the thin filaments and serving as an anchor to which the thin filaments
are attached.

 The A band too is bisected by a thin line, the M line or M disk.

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 The entire contractile unit, consisting of bundles of thick filaments interleaved
at either end with bundles of thin filaments, is called the sarcomere.

 This arrangement allows the thick and thin filaments to slide past each other
causing a progressive shortening of each sarcomere.

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How does myosin move?
 The interaction between actin and myosin, like that between
all proteins and ligands, involves weak bonds.

 When ATP is not bound to myosin, a face on the myosin head

group binds tightly to actin.

 When ATP binds to myosin and is hydrolyzed to ADP and

phosphate, a coordinated and cyclic series of conformational
changes occurs in which myosin releases the F-actin subunit
and binds another subunit farther along the thin filament.

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The cycle has four major steps:

 In step (1), ATP binds to myosin and a cleft

in the myosin molecule opens, disrupting the
actin-myosin interaction so that the bound
actin is released.

 ATP is then hydrolyzed in step (2), causing a

conformational change in the protein to a
“high-energy” state that moves the myosin head
and changes its orientation in relation to the
actin thin filament.

 Myosin then binds weakly to an F-actin

subunit closer to the Z disk than the one
just released.

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 As the phosphate product of ATP hydrolysis is
released from myosin in step (3), another
conformational change occurs in which the
myosin cleft closes, strengthening the
myosin-actin binding.

 This is followed quickly by step (4), a “power

stroke” during which the conformation of the
myosin head returns to the original resting
state, pulling the actin filament over itself
towards the center of the sarcomere. ADP is
then released to complete the cycle.

 The thick filament thus actively slides forward past the

adjacent thin filaments.

 This process, coordinated among the many

sarcomeres in a muscle fiber, brings about
muscle contraction.

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 The interaction between actin and myosin must be regulated so that
contraction occurs only in response to appropriate signals from the nervous
system.

 Tropomyosin binds to the thin filament, blocking the attachment sites for
the myosin head groups.

 When a muscle is stimulated by a nerve impulse, it causes release of Ca2+

from the sarcoplasmic reticulum.

 The released Ca2+ binds to troponin-C which in turn interacts with troponin-
I and reverses the inhibitory effect on actin-myosin interaction.

 Actin is switched on when tropomyosin moves to a new position on each

actin molecule so that the myosin-binding sites are exposed.

 Contraction follows.

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 When the excitation ceases, Ca2+ ions are actively pumped back by an
ATP-driven calcium pump.

 Ca2+ ion concentration soon decreases below the threshold for

contractile activity and relaxation of sarcomere begins.

 Tropomyosin-troponin complex inhibits the ATPase activity, cross-

bridges are broken, actin and possibly myosin are “switched off” and
the sarcomere reverts to its normal resting state

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RIGOR MORTIS
 Rigor mortis is the stiffening of the body after death because of a loss of ATP
from the muscles.

 In normal muscle, concentration of ATP remains fairly constant even in

strenuous activity because of increased metabolic activity and the actions of
creatine phosphokinase and adenylate kinase.

 Creatine phosphokinase catalyzes transfer of phosphate from

phosphocreatine to ADP.

 Adenylate kinase provides additional ATP by catalyzing the reaction

2ADP ATP + AMP

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 Effects of ATP depletion are:

1. Intracellular Ca2+ concentration is no longer controlled and

2. Myosin will exist exclusively in the myosin-ADP complex and bound to actin,
since ATP is required for dissociation of the actin-myosin complex.

 Rigor mortis begins throughout the body at the same time but the smaller
muscles such as facial, neck, arm and shoulders are affected first.

 Rigor normally appears around three hours after death has occurred.

 Peak of rigor mortis appears approximately 12 hours after death.

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MICROFILAMENT & MICROTUBULES
 Microfilaments and microtubules are key components of
the cytoskeleton in eukaryotic cells.

 Together they allow the cell to hold its shape, and move itself and its
organelles.

 Individual subunits of microfilaments are known as globular actin (G-actin).

G-actin subunits assemble into long filamentous polymers called F-actin. 

 Two parallel F-actin strands form the double helix structure of

microfilaments.

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FUNCTION OF MICROFILAMENT

 Microfilaments form the dynamic cytoskeleton, which gives structural support

to cells and links the interior of the cell with the external environment.

 Microfilaments provide cell motility. e.g., Filopodia, Lamellipodia.

 During mitosis, intracellular organelles are transported by motor proteins to

the daughter cells along actin cables.

 In muscle cells, actin filaments are aligned and myosin proteins generate forces
on the filaments to support muscle contraction.

 In non-muscle cells, actin filaments form a track system for cargo transport
that is powered by non-conventional myosins such as myosin V and VI. Non-
conventional myosins use the energy from ATP hydrolysis to transport cargo
(such as vesicles and organelles) at rates much faster than diffusion.

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 Microtubules are composed of globular proteins called tubulin.
They form heterodimers of alpha and beta tubulin. 

FUNCTIONS OF MICROTUBULES

 Microtubules determine the cell structure.

 Microtubules form the spindle apparatus to divide the chromosome

directly during cell division (mitosis).

 Microtubules provide transport mechanism for vesicles containing

essential materials to the rest of the cell.

 They form a rigid internal core that is used by microtubule-

associated motor proteins (MAPs) such as Kinesin and Dyenin to
generate force and movement in motile structures such as cilia and
flagella.

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