2H NMR study of molecular motion in collagen fibrils

Jelinski, Lynn W.; Sullivan, C. E.; Torchia, Dennis A.

doi:10.1038/284531a0

Cited by 98 publications

(87 citation statements)

References 17 publications

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“…In contrast, the liquid-crystalline model for collagen (35) suggests a significant degree of disorder, especially in the collagen fiber gap regions. In addition, nuclear magnetic resonance studies indicate a considerable degree of rotational freedom for individual molecules in the collagen fiber (36). The distribution of binding sites for SPARC, DDR2, and VWF along the collagen circumference supports the notion of considerable flexibility within the collagen fibril.…”

Section: Resultsmentioning

confidence: 74%

Implications for collagen I chain registry from the structure of the collagen von Willebrand factor A3 domain complex

Brondijk

Bihan²,

Farndale³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Fibrillar collagens, the most abundant proteins in the vertebrate body, are involved in a plethora of biological interactions. Plasma protein von Willebrand factor (VWF) mediates adhesion of blood platelets to fibrillar collagen types I, II, and III, which is essential for normal haemostasis. High affinity VWF-binding sequences have been identified in the homotrimeric collagen types II and III, however, it is unclear how VWF recognizes the heterotrimeric collagen type I, the superstructure of which is unknown. Here we present the crystal structure of VWF domain A3 bound to a collagen type III-derived homotrimeric peptide. Our structure reveals that VWF-A3 interacts with all three collagen chains and binds through conformational selection to a sequence that is one triplet longer than was previously appreciated from platelet and VWF binding studies. The VWF-binding site overlaps those of SPARC (also known as osteonectin) and discodin domain receptor 2, but is more extended and shifted toward the collagen amino terminus. The observed collagen-binding mode of VWF-A3 provides direct structural constraints on collagen I chain registry. A VWF-binding site can be generated from the sequences RGQAGVMF, present in the two α1 (I) chains, and RGEOGNIGF, in the unique α2(I) chain, provided that α2(I) is in the middle or trailing position. Combining these data with previous structural data on integrin binding to collagen yields strong support for the trailing position of the α2(I) chain, shedding light on the fundamental and long-standing question of the collagen I chain registry.X-ray crystallography | extracellular matrix V on Willebrand factor (VWF) mediates blood platelet adhesion to sites of vascular damage and is essential for platelet adhesion under conditions of high shear stress. Defective processing and/or mutations in VWF are associated with various haemostatic disorders such as haemolytic uremic syndrome, thrombotic thrombocytopenic purpura, and von Willebrand disease, which is the most common genetic bleeding disorder in humans (1). The mature VWF protein consists of 2,050 amino acid monomers that are disulfide-linked into multimers ranging from 0.5 to 10 MDa in size with the larger multimers being more active in hemostasis. VWF multimer size is regulated by ADAMTS13 through proteolytic cleavage within the VWF A2 domain. Upon vascular damage, the VWF A3 domain interacts with exposed collagens I and III. Subsequent transient interactions between the VWF A1 domain and Glycoprotein Ibα (GPIbα) on the surface of blood platelets reduces platelet velocity to allow subsequent stable platelet adhesion and activation, which ultimately results in thrombus deposition.The crystal structure of the collagen-binding VWF A3 domain has been solved (2-4) and the collagen-binding site within the A3 domain has been mapped in extensive site-directed mutagenesis studies and by NMR (5-7). However, detailed structural information on the VWF-collagen interaction has thus far been lacking.Collagens, the most abundant proteins in mammalian...

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Section: Resultsmentioning

confidence: 74%

Implications for collagen I chain registry from the structure of the collagen von Willebrand factor A3 domain complex

Brondijk

Bihan²,

Farndale³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Previous 2H and 13C NMR studies of labeled collagen fibrils have shown that rapid, r-=10-s, reorientation of the peptide backbone occurs about the long axis ofthe helix through an angle of ca. 300 (1)(2)(3)(4). These studies also have provided strong evidence that the amino acid side chains exhibit extensive internal motions in the fibrils (3,5 (9) that accounts for spectral distortion due to a finite pulse width.…”

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confidence: 74%

Characterization of leucine side-chain reorientation in collagen-fibrils by solid-state 2H NMR.

Batchelder¹,

Sullivan²,

Jelinski³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

109

135

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We have used 2H quadrupole-echo NMR spectroscopy to study the molecular dynamics of the leucine side chain in collagen fibrils labeled with [2H10]leucine. X-ray crystallographic studies ofleucine and small leucyl-containing peptides and proteins [Benedetti, C. (1977) at 313 nm to establish purity. The percentage incorporation of deuterium (43%) was determined by chemical ionization gas chromatography/mass spectrometry of N-acetylmethyl ester derivatives of the hydrolyzed protein. Radiotracer (3H and 14C) analysis showed that leucine was the only amino acid labeled during biosynthesis.2H NMR spectra were observed for polycrystalline D,L-[2H10]leucine over the temperature range -45°C to + 100°C (Fig. 2). Because of the size ofthe methylene and methine deuteron quadrupolar splitting, Avq(120 kHz), only a part of the spectrum arising from these deuterons is shown. Rapid threefold rotation of the methyl groups in leucine averages Avq to 38 kHz at -45°C, and these six deuterons display the classic axially symmetric quadrupolar powder pattern. Over the temperature range studied, very little additional side chain motion

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“…Line shape analysis has most widely been used in 2 H NMR for various reasons: i) spectral interpreta-tion is facilitated since the 2 H gradient field tensor is almost perfectly axially symmetric for most CD bonds and its unique principal axis aligned with the CD vector within 38 (59,120); ii) quadrupole splittings of 2 H are about 200 kHz (116,121) and spectra are therefore sensitive to motions with correlation times shorter than the ms; iii) finally, in addition to cell culture specific labeling, fibers can sometimes be 2 H-labeled by extensive soaking in D 2 O (19, 116). 2 H spectra are recorded using the solid echo technique to correct phasing and baseline problems due to receiver dead time (122).…”

Section: Signal Intensity Line Width and Line Shape Analysismentioning

confidence: 99%

“…First, 2 H NMR spectra of alanine-labeled collagen were analyzed by Jelinski et al (120) according to a model in which the fiber backbone undergoes rapid reorientation about the long axis of the molecule. By fitting the experimental spectra with calculated spectra based on a two-site jump model, the reorientation angle was determined to be of $308 (120,123) and the average angle between the alanine C a -C b bond axis and long axis of the collagen helix was estimated to be $758 (123). In addition, the motion of leucine side chains in collagen was also evidenced by 2 H NMR and could be explained by a fast exchange between two predominant conformations (123,124).…”

Section: Signal Intensity Line Width and Line Shape Analysismentioning

confidence: 99%

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

Arnold

Marcotte

2009

Concepts Magnetic Resonance

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As a consequence of evolutionary pressure, various organisms have developed structural fibers displaying a range of exceptional mechanical properties adapted specifically to their functions. An understanding of these properties at the molecular level requires a detailed description of local structure, orientation with respect to the fiber and size of constitutive units, and dynamics on various timescales. The size and lack of long-range order in these protein systems constitute an important challenge to classical structural techniques such as high-resolution NMR and X-ray diffraction. Solidstate NMR overcomes these constraints and is uniquely suited to the study of these inherently disordered systems. Solid-state NMR experiments developed or applied to determine structure, orientation, and dynamics of these complex proteins will be reviewed and illustrated through examples of their applications to fibers such as spider and silkworm silks, collagen, elastin, and keratin.

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2H NMR study of molecular motion in collagen fibrils

Cited by 98 publications

References 17 publications

Implications for collagen I chain registry from the structure of the collagen von Willebrand factor A3 domain complex

Implications for collagen I chain registry from the structure of the collagen von Willebrand factor A3 domain complex

Characterization of leucine side-chain reorientation in collagen-fibrils by solid-state 2H NMR.

Studying natural structural protein fibers by solid‐state nuclear magnetic resonance

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