Received: 12 June 2020 Revised: 28 July 2020 Accepted: 30 July 2020

DOI: 10.1002/mrc.5084

RESEARCH ARTICLE

Probing short and long-range interactions in native

collagen inside the bone matrix by BioSolids CryoProbe

Nidhi Tiwari1,2 | Sebastian Wegner3 | Alia Hassan3 | Navneet Dwivedi1,4 |

RamaNand Rai2 | Neeraj Sinha1

1
Centre of Biomedical Research,
SGPGIMS Campus, Lucknow, 226014,
Abstract
India Solid-state nuclear magnetic resonance is a promising technique to probe bone
2
Department of Chemistry, Institute of mineralization and interaction of collagen protein in the native state. However,
Sciences, Banaras Hindu University,
many of the developments are hampered due to the low sensitivity of the
Varanasi, 221005, India
3 technique. In this article, we report solid-state nuclear magnetic resonance
Bruker BioSpin Corporation, Fällanden,
Switzerland (NMR) experiments using the newly developed BioSolids CryoProbe™ to
4
Department of Physics, Integral access its applicability for elucidating the atomic-level structural details of
University, Lucknow, 226026, India collagen protein in native state inside the bone. We report here approximately
Correspondence a fourfold sensitivity enhancement in the natural abundance 13C spectrum
Neeraj Sinha, Centre of Biomedical compared with the room temperature conventional solid-state NMR probe.
Research, SGPGIMS Campus, Raebarelly
With the advantage of sensitivity enhancement, we have been able to perform
Road. Lucknow 226014, India.
Email: neerajcbmr@gmail.com; natural abundance 15N cross-polarization magic angle spinning (CPMAS) and
1
neeraj.sinha@cbmr.res.in two-dimensional (2D) H–13C heteronuclear correlation (HETCOR)

Funding information
Science and Engineering Research Board,
Grant/Award Number:
EMR/2015/001758; Department of Science
and Technology (DST); Council of
Scientific and Industrial Research (CSIR);
SERB, Grant/Award Number:
EMR/2015/001758
experiments of native collagen within a reasonable timeframe. Due to high
sensitivity, 2D 1H/13C HETCOR experiments have helped in detecting several
short and long-range interactions of native collagen assembly, thus signifi-
cantly expanding the scope of the method to such challenging biomaterials.
KEYWORDS
bone, collagen, CryoProbe, magic angle spinning, solid-state NMR

1 | INTRODUCTION functional behavior of collagen, it has always been the

Collagen is a ubiquitous and predominant protein found
in mammals and contributes to one-third of the whole
body protein.[1] It is a structural component of the
extracellular matrix (ECM) of various connective tissue,
such as bone and cartilage, as well as skin and basement
membranes.[2] The primary function of the triple-helical
collagen is to provide strength and flexibility in bone
and ligaments on exposure to external stresses.[3] In
addition to this, collagen proteins are essential for a
broad range of functions, including entrapment, local
storage, delivery of growth factors, angiogenesis, tissue
morphogenesis, and tissue repair.[2,4] Due to the broad
utmost important research area. The 90% of the organic
part in bone ECM is made up of Type I collagen, respon-
sible for the mechanical stability and flexibility of the
bone. Additionally, collagen plays a predominant role in
the bone bio-mineralization process by affecting the
nucleation, growth, structure, and orientation of bone
appetite.[5] The unique triple-helical structure of collagen
consists of an assembly of three parallel helices with the
tripeptide repeating sequence of Gly-Xaa-Yaa, where
glycine (Gly) occupies every third position and buried
inside the center of the triple-helical strands, and X, Y
positions filled by mostly proline (Pro) (28%
abundance) and hydroxyproline (Hyp) (38%

Magn Reson Chem. 2020;1–9. wileyonlinelibrary.com/journal/mrc © 2020 John Wiley & Sons, Ltd. 1
2 TIWARI ET AL.
abundance) are exposed towards outside.[6–8] The
interchain hydrogen bonding holds the triple-helical col-
lagen assembly while Hyp confers the additional stability
of the collagen protein through water-mediated hydrogen
bonding and stereoelectronic effects.[9,10] Most of the
structural studies of Collagen protein confined around
the model peptides,[7] extracted collagen,[11] and molecu-
lar dynamics simulations.[12] Different spectroscopic
techniques X-ray,[13,14] Fourier transformed infrared
(FT-IR),[15] Raman spectroscopy,[16–18] micro-computed
tomography (CT),[15] and atomic force microscopy
(AFM)[19] give the structural details of static state but fail
to detect the dynamics and orientation-dependent molec-
ular interaction in a native environment. Inside the bone,
the native environment of collagen is formed by deposi-
tion of mineral, lipids, noncollagenous proteins (NCPs),
citrate as well as water, and its associated interaction with
them.[20,21] All these interactions present inside the bone
are responsible for the different structural arrangement of
native collagen protein than the extracted form of colla-
gen.[22] Understanding the structural and functional
behavior of collagen triple helix assembly in its actual
native state in healthy and diseased conditions will pave
the way for the development of treatment methods for
various bone degenerative diseases,[23] tissue engineering
methods of artificial cartilage,[24] and bone implants.[25]
Solid-state nuclear magnetic resonance (NMR) is a
robust technique to probe structural and dynamical
details along with associated interaction among
different components of such complex heterogeneous
biomaterial.[26–31] Still, it has limited applicability in cap-
turing the detailed structural framework of native colla-
gen due to the low concentration of organic components
inside the bone. Natural abundance spectrum of collagen
suffers from weak signal-to-noise (S/N) ratio because of
low sensitivity and limited isotopic concentration. Due to
low sensitivity, it is difficult to explore the various inter-
actions present inside the biological system at the molec-
ular and atomic level that is beyond the detectable
range of solid-state NMR. Sensitivity enhancement is a
significant concern that has to be overcome to get high-
resolution structural details, along with associated
interaction among different components of bone in
natural abundance in a limited time framework. One
powerful approach for boosting the sensitivity of the
natural abundance spectrum is high-field dynamic
nuclear polarization (DNP)-based solid-state NMR exper-
iments that have become feasible with the development
of high-power microwave sources and cryogenic MAS
probes.[32–36] Use of BioSolids CryoProbe is another
recently developed approach to improve the sensitivity
without line broadening. Cryogenically cooling the RF
coil and preamplifier electronics leads to a boost in S/N
by 3–4 folds and hence faster data acquisition by at least
an order of magnitude while keeping the sample in its
original composition along with its natural line width.[37]
In the present article, we report the advantage of sen-
sitivity enhancement achieved in natural abundance 13C
and 15N spectra of native collagen inside the bone matrix
by using an HCN cross-polarization magic angle spinning
(CPMAS) CryoProbe for solid-state NMR. Due to sensitiv-
ity enhancement, it allows recording high-resolution
two-dimensional (2D) 1H–13C heteronuclear correlation
(HETCOR) experiments of collagen in natural isotopic
abundance within a limited timeframe. With the help of
high-resolution 2D 1H–13C HETCOR spectra recorded at
short and long contact times, we can probe the short and
long-range interactions within the spatial proximity of
collagen inside the bone matrix in its actual native envi-
ronments. These interactions play a predominant role in
the structural stability and functional mechanism of
collagen inside the bone matrix.
2 | E XP E R I M EN T A L S E C T I O N
2.1 | Sample preparation
All the solid-state NMR experiments were performed on
the cortical femora bone of goat (Capra hircus, 2–3 years
old). Small size flakes were obtained by filing the intact
bone with the help of a scalpel. These flakes had the
same morphology as that of intact bone. The bone
sample was not subjected to any kind of pretreatment or
processing because it causes the alteration in the homo-
geneity and water content of the bone matrix.[38]
2.2 | Solid-state NMR experimental
parameter
All solid-state NMR spectra were collected on a 14.1 T
narrow bore Bruker AVANCE NEO spectrometer operat-
ing at 600.3 MHz (1H), 150.9 MHz (13C), and 60.8 MHz
(15N) Larmor frequencies. This spectrometer was
equipped with a cryogenically cooled 3.2-mm triple-
resonance 1H/13C/15N CPMAS probe (BioSolids Cryo-
Probe). The magic angle spinning (MAS) speed was set to
12 kHz using Bruker's MAS3 pneumatic unit within an
accuracy of ±2 Hz. 1H π/2 pulse was calibrated to 2.50 μs.
For 13C CPMAS spectrum, ramp cross-polarization
sequence at Hartman Hahn match condition with spinal-
64[39] 1H decoupling of 52.08 kHz and 1.25-ms contact
time was set. The 1D 13C spectrum was acquired with
512 scans at 5-s recycle delay. The 15N CPMAS spectrum
was recorded with 878 scans at 3-s recycle delay using
TIWARI ET AL. 3

cross-polarization sequence with spinal-64 1H decoupling
of 50 kHz and 1.0-ms contact time. The 2D 1H–13C
CP-MAS HETCOR experiments were acquired using
frequency-switched Lee-Goldberg (FSLG)[40,41] 1H homo-
nuclear decoupling. The 256 t1 increments for a total
acquisition of 5.36 ms with 16 scans and 3-s recycle delay
were used for recording the experiment. 2D 1H–13C het-
eronuclear correlation (FSLG-HETCOR) experiments
were carried out at 100 and 500-μs contact times.
four folds (S/N 156:1) in the natural abundance 1H–13C
CPMAS spectrum of native collagen (Figure 1 and
Table S1). We have assigned most of the 13C resonances
from an organic matrix. Resonances of Gly, Hyp, and Pro
from the backbone of collagen are similar to the spec-
trum recorded with the conventional solid-state NMR
method (Figure 2).[38,45] The signal intensity of ring
vcarbon resonances of aromatic amino acids (AAAs) such
as Phe, Tyr, Trp, and histidine (His) resonances of colla-
gen protein around 129 ppm is significantly enhanced

3 | R E S UL T S A ND D I S CUS S I O NS

The physiochemical properties of a bone microstructure

are defined by the associated intermolecular and intra-
molecular interaction among the various constituents.
The triple-helical structure of collagen has frequent inter-
actions involving hydration network and hydroxyproline
that endorse the nonspecific self-association,[2] while
charged and hydrophobic interactions confer specificity
and a high affinity for self-association.[42–44] Probing
these interactions responsible for peculiarity and self-
association of the triple-helical structure of collagen to
the higher order structure would be helpful in developing
a better understanding of the structural and functional
role of collagen in bone microstructure. In this article,
intrastrand, intrahelix, and interhelix interactions present
in the collagen triple-helical structure inside the bone are
probed by 1H–13C HETCOR experiments at short and
long contact times.
FIGURE 2 Resonance assignment of 1H–13C cross-
polarization (CP) magic angle spinning (MAS) spectra of collagen
3.1 | Natural abundance 13C CPMAS in natural isotopic abundance recorded at 600-MHz solid-state
spectrum of native collagen nuclear magnetic resonance (NMR) spectrometer equipped with a
3.2-mm triple resonance 1H/13C/15N cross-polarization MAS
Probing the organic part of the bone with a BioSolids (CPMAS) BioSolids CryoProbe with approximately fourfold
CryoProbe causes an absolute increase in sensitivity by sensitivity enhancement

1
FIGURE 1 H–13C cross-polarization (CP) magic angle spinning (MAS) spectra of native collagen recorded at (a) 600-MHz solid-state
nuclear magnetic resonance (NMR) equipped with a 3.2-mm Direct Variable Temperature (DVT) probe. (b) 600-MHz solid-state NMR
spectrometer equipped with a 3.2-mm triple resonance 1H/13C/15N cross-polarization MAS (CPMAS) BioSolids CryoProbe
4 TIWARI ET AL.
(Figure 2). Moreover, we have observed resonances from
citrates at 74–76 ppm, which is closer to the inorganic
surface. Due to low abundance, resonances from ring
carbons and citrate show very weak intensity in conven-
tional solid-state NMR spectra even with a very high
signal averaging. The increased signal intensity of the
aromatic ring carbon resonances observed with Cryo-
Probe opens up the possibility to probe various hydro-
phobic interactions present inside the collagen triple
helices in its native environment.
3.2 | Natural abundance 1H–15N CPMAS
spectrum of native collagen
Acquiring 15N spectra of collagen protein in natural
abundance is quite challenging because of very low nat-
ural occurrence (0.37%) and low gyromagnetic ratio
(−2.7126 × 107 rad T − 1 s − 1) of 15N.[46] By reducing
1
H T1 accompanied by a long signal averaging time, we
can detect 15N resonances in natural abundance. Ear-
lier, the spectrum acquired by sensitivity enhancement
with paramagnetic doping of Cu (II) EDTA in the
sample[47–49] suffers from line-broadening. With the
help of a BioSolids CryoProbe, we could obtain a high-
resolution natural abundance 1H–15N CPMAS spectrum
with a significant increase in sensitivity (S/N 42:1)
along with faster acquisition without any paramagnetic
doping (Figure 3). We have assigned the resonances
originating exclusively from the Pro/Hyp backbone at
121.6 ppm, Gly at 108 ppm,[50] and 15N side chains of
histidine and arginine.[47] Resonances from arginine
(84.4 ppm and 70.8 ppm) and lysine (32 ppm) were also
assigned,[51] which were not possible to detect in the
conventional solid-state NMR CPMAS spectrum of 15N
(Figure 3).
3.3 | Natural abundance 2D 1H–13C FSLG-
HETCOR solid-state NMR
To probe the interhelix and intrahelix molecular interac-
tions in the spatial proximity of collagen in its actual
native environment, we performed 2D 1H − 13C heter-
onuclear correlation (FSLG-HETCOR) experiments at
100 and 500 μs contact times (Figure 4). Well-resolved 2D
1
H − 13C HETCOR spectra of native collagen were
acquired in a reasonable timeframe. FSLG-HETCOR
experiment performed at 100-μs contact time shows a
one-bond C − H ( 1.0–1.5 Å) correlation. In compari-
son, at 500 μs contact time, we get the long-range
( 2–6 Å) correlation within the spatial proximity of col-
lagen inside the bone matrix, as shown in Figure 4. The
appearance of the additional peak in the carbonyl region,
aromatic region, and NH region in the 2D 1H–13C
HETCOR spectrum of collagen indicated the presence of
a long-range correlation. Moreover, with the advantage
of increased sensitivity, we could explicitly assign the car-
bonyl group of different residues of collagen protein,
probe the various molecular interactions, and detect
the interaction of ring carbons of AAAs with different
residues of collagen protein.
FIGURE 3 Resonance assignment of
1
H–15N cross-polarization (CP) magic angle
spinning (MAS) spectra of collagen in natural
isotopic abundance recorded at 600-MHz solid-
state nuclear magnetic resonance (NMR)
spectrometer equipped with a 3.2-mm triple
resonance 1H/13C/15N cross-polarization MAS
(CPMAS) BioSolids CryoProbe with significant
sensitivity enhancement
TIWARI ET AL.
FIGURE 4 Overlay of two-dimensional
1
H–13C frequency-switched Lee-Goldberg
heteronuclear correlation (FSLG-HETCOR) of
native collagen at 500-μs (green) and 100-μs
(pink) correlation times
3.3.1 | Short and long-range interaction in
collagen
The 2D 1H–13C HETCOR experiments provide atomic-
level piercing insights into the assembly of collagen triple
helix inside the bone. The overlay of 2D 1H–13C
HETCOR spectra recorded at 100 μs and 500 μs is shown
in Figure 4. At 100 μs, one bond intramolecular interac-
tion could be seen (Figure S1) in triple-helical collagen
protein. Dotted lines represent correlations among Hyp
Cγ, Hyp Cβ, and Hyp C. Interaction between Pro Cβ and
Pro Cγ also shows one bond correlation. Resonances
from carbonyl group, NH group, and ring carbons of aro-
matic groups of collagen protein could not be observed
with 100-μs contact time. Additionally, resonances from
Ala Cβ are not observable due to limited magnetization
transfer up to one bond. 2D 1H–13C HETCOR spectra
recorded at 500-μs contact time allows magnetization
transfer up to 6-Å distances enabling us to explore the
spatial proximity of collagen triple helix. Long-range
intrahelix and interhelix interactions could be seen
among the various residues of collagen protein, as shown
in Figures 5a and 6a. Ring carbon of Hyp show correla-
tion with ring carbon of Pro indicating the presence of
hydrophobic interactions. These interactions are present
5
in interhelix collagen assembly confirmed by depicting
the closest distance in the crystal structure of collagen
protein (Protein Data Bank (PDB) ID: 1Q7D), as shown
in Figure 6b(iii). A long-range intrahelix interactions can
be seen between Gly Cα and Pro Cβ of collagen protein
Figure 5b(iii), because these residues engaged in the
hydrogen bonding network, which is responsible for the
stability of the triple-helical structure of collagen protein.
The molecular interactions between Arg Cα (arginine)-
Asp (aspartic) Cα and Lys (lysine) Cε-Asp Cβ suggests
the possibility of cation-pair interactions that had been
studied in collagen peptide.[42] A molecular interaction is
shown by distance measurement between Arg (basic) and
Glu (acid) in Figure 5(iv).
Correlation of carbonyl resonance of collagen
As observed in the 1H–13C HETCOR spectra (Figure 5a)
of collagen protein, it could be seen that the carbonyl
regions (168–176 ppm) distinctly arise from highly abun-
dant residues (Hyp, Pro, and Gly) of native collagen.
Figure 5a shows the correlation between the carbonyl
carbon and the cyclic proton of Hyp, and Pro represented
with the dotted lines. Additionally, the correlation
between the NH proton of glycine and the carbonyl
group confirms the presence of interchain N–H (Gly)



6 TIWARI ET AL.
O=C (Xaa) hydrogen bonds that are responsible for the
alignment of three chains in the collagen triple helix.[7]
In collagen triple helix, the correlation among Cα protons
of Hyp, Pro, and Gly and carbonyl carbon also suggest
the presence of Cα–H(Gly/Yaa)


O=C(Xaa/Gly) type of
hydrogen bonds as reported by Bella et al.[52] These
interchain Cα–H–O=C hydrogen bonds in collagen
protein are responsible for a large ensemble of weak
interactions in the inner core of the triple helix, add
extra stability to the structure of collagen protein
(Figure 5a(ii)).
Correlation of aromatic resonances of collagen
Aromatic amino acid residues are relatively rare in the
triple-helical assembly of collagen. Phenylalanine (Phe)
FIGURE 5 (a) Two-dimensional
1
H–13C frequency-switched Lee-
Goldberg heteronuclear correlation
(FSLG-HETCOR) spectrum of native
collagen at 500 μs showing correlation
of carbonyl resonances with Hyp, Pro,
and Gly residues, and other molecular
interactions. (b) (i) One collagen triple
helix in the crystal structure of integrin-
binding collagen PDB ID: 1Q7D.
(ii) Intrahelix hydrogen bonding
network among Gly, Pro, and Hyp
residues. Expanded view of intrahelix
molecular interactions are indicated by
depicting the distance between (iii) Gly-
Pro (hydrogen bonding). (iv) Glu-Arg
(charge pair interactions). (v) NH-π
interactions between NH proton and
ring carbons of phenylalanine (Phe)
and tyrosine (Tyr) AAAs constitute 1.4% and 0.3% of total
amino acid present in the collagen of the mammalian
bone.[53] These AAAs have a very specialized role in
higher order structure and function of collagen protein.
They appear to play a key role in accelerating the aggre-
gation of the triple-helical structure of collagen and
affecting their biological activity.[44] Earlier, a DNP-based
solid-state NMR experimental study of collagen protein
of bone provides evidence for CH-π interaction between
Phe and Pro/Hyp of neighboring asymmetric pair of tri-
ple helix in collagen assembly.[44,54] Figure 6b(ii) shows
the molecular interactions of Phe with Pro/Hyp in the
closest neighboring helix. Here, we observe the molecular
interactions of aromatic residues with other interstrand
and intrastrand residues of collagen triple helix.
TIWARI ET AL. 7

FIGURE 6 (a) Two-dimensional

1
H–13C frequency-switched
Lee-Goldberg heteronuclear correlation
(FSLG-HETCOR) spectrum of native
collagen at 500-μs contact time showing
correlation of aromatic resonances with
NH protons of Lys, Gly, and Asp
residues along with other molecular
interactions. (b) (i) Interhelix molecular
interactions in asymmetric pairs of
collagen triple helix in the crystal
structure of integrin-binding collagen
PDB ID: 1Q7D. Expanded view of
interhelix molecular interactions are
shown by depicting the distance
between (ii) Phe-Pro-Hyp (CH- π
interactions). (iii) Hyp-Pro (hydrophobic
interactions)

As shown in Figure 6a, the 1H spectral region of
6–9 ppm is assigned to NH proton of collagen protein.
Resonances observed at 124–132 ppm/6.5–8.5 ppm
(13C/1H) are assigned to ring carbons and NH protons of
Tyr, Phe, and His residues of collagen protein. The corre-
lation between NH protons of Lys/Gly and Asp and ring
carbon of aromatic residues shows the intrahelix and
interhelix polarization transfer through space in collagen
assembly. Additionally, the cross peak observed at
124–132 ppm/6.5–8.5 ppm (13C/1H) indicates the polari-
zation transfer from NH proton to ring carbon of AAAs;
Figure 5b(v) shows the measurement of the distance
between NH proton and ring carbon of Phe in the struc-
ture (PDB ID 1Q7D). Interactions between Lys (Cζ/Ηζ)
42.1/7.82 ppm with ring carbon of AAAs indicate the
presence of polar NH-π/Cation-π) hydrophobic interac-
tions. These hydrophobic interactions (CH/NH-π, cation-
π) are responsible for local or global stability of collagen
protein.[43,55,56]
The atomic-level investigation of the multiscale het-
erogeneous structure of bone has been challenging due to
poor spectral resolution and sensitivity. However, MAS-
DNP solid-state NMR methods are highly efficient in sen-
sitivity enhancement for elucidating the structural details
of the biological system, such as bone at natural isotopic
abundance.[51,57] Despite the advantage of sensitivity
enhancement, due to the requirement of the low-temper-
ature, molecular motion is significantly slowed down,
8 TIWARI ET AL.
and molecules are trapped in a range of conformations
that leads to slightly different chemical shifts along with
the broadening of the signals and a reduction in the
resolution. Moreover, with the use of stable biradical for
doping the sample, DNP-enhanced ssNMR cannot be
considered a noninvasive technique.[36] A recent develop-
ment in multidimensional 1H detected NMR experiments
had been employed to study the bone microstructure.[58]
1
H-detected NMR measurements on the bone under
ultrafast MAS (>60 kHz) conditions led to an improve-
ment in spectral resolution and sensitivity enhancement
by effective suppression of 1H–1H homonuclear dipolar
coupling and increased transverse relaxation time.
Hence, 1H detected NMR experiments enables the
manipulation of long-lived magnetization for potential
applications. The proton-detected HETCOR experiment
and low-γ nuclei homonuclear correlation achieved by
transfer of polarization could offer significant insights
into molecular structures and dynamics.[59] A develop-
ment in the BioSolids CryoProbe is another significant
advancement in solid-state NMR spectroscopy, facilitat-
ing to get 3–4 fold sensitivity enhancement. The MAS
speed in BioSolids CryoProbe is limited up to 20 kHz. In
the future, multidimensional 1H detected solid-state
NMR experiments using BioSolids CryoProbe (noninva-
sive) would be more advantageous in elucidating the
atomic-scale structure along with the dynamics of the
biological system. Hence, all these developments such as
proton-detected solid-state NMR experiments, CPMAS
BioSolids CryoProbe,[37] and MAS-DNP-based solid-state
NMR experiments can complement each other to get
structural and dynamical details of biological systems in
natural abundance.
4 | C ON C L US I ON
We observed approximately four-fold sensitivity enhanced
high-resolution natural abundance 13C CPMAS spectra of
collagen inside the bone acquired with recently developed
BioSolids CryoProbe. Additionally, we could detect the
natural abundance of 15N resonances of collagen protein
in its native state with reasonable signal averaging. With
the advantage of sensitivity enhancement, we could
observe long-range intrahelix and interhelix, along with
intrastrand molecular interactions in the spatial proximity
of native collagen through 2D 1H–13C HETCOR experi-
ments. The boost in sensitivity by using the BioSolids
CryoProbe enables us to examine the noncovalent interac-
tions such as hydrogen bonds, electrostatic interaction, as
well as hydrophobic interactions responsible for folding
stability of collagen protein. Understanding ultrastructural
details and associated interactions present in the native
environment of collagen inside bone would be useful in
comprehending its structural and functional mechanism
during bone homeostasis and helpful in developing pre-
vention and care therapy for the diseases related to colla-
gen structural disorder.
A C KN O WL ED G EME N T S
The authors gratefully acknowledge SERB
(EMR/2015/001758), the Council of Scientific and Indus-
trial Research (CSIR), and the Department of Science
and Technology (DST), India, for financial support.
ORCID
Neeraj Sinha https://orcid.org/0000-0003-3235-6127
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SU PP O R TI N G I N F O RMA TI O N
Additional supporting information may be found online
in the Supporting Information section at the end of this
article.
How to cite this article: Tiwari N, Wegner S,
Hassan A, Dwivedi N, Rai RN, Sinha N. Probing
short and long-range interactions in native
collagen inside the bone matrix by BioSolids
CryoProbe. Magn Reson Chem. 2020;1–9. https://
doi.org/10.1002/mrc.5084