NMR Studies Demonstrate a Unique AAB Composition and Chain Register for a Heterotrimeric Type IV Collagen Model Peptide Containing a Natural Interruption Site

Xiao, Jianxi; Sun, Xiuxia; Madhan, Balaraman; Brodsky, Barbara; Baum, Jean

doi:10.1074/jbc.m115.654871

Cited by 23 publications

(24 citation statements)

References 46 publications

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“…These peptide studies have greatly advanced our understanding of the roles of Gly mutations in triple helix stability, conformation, and folding. Meanwhile, peptide models of natural interruptions have also displayed localized alterations in conformation . However, the molecular basis underlying the contradictory functional consequences of Gly mutations and natural interruptions is still poorly understood.…”

Section: Introductionmentioning

confidence: 99%

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CD and NMR investigation of collagen peptides mimicking a pathological Gly-Ser mutation and a natural interruption in a similar highly charged sequence context

Sun

Liu

et al. 2015

Protein Science

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Even a single Gly substitution in the triple helix domain of collagen leads to pathological conditions while natural interruptions are suggested to play important functional roles. Two peptides-one mimicking a pathological Gly-Ser substitution (ERSEQ) and the other one modeling a similar natural interruption sequence (DRSER)-are designed to facilitate the comparison for elucidating the molecular basis of their different biological roles. CD and NMR investigation of peptide ERSEQ indicates a reduction of the thermal stability and disruption of hydrogen bonding at the Ser mutation site, providing a structural basis of the OI disease resulting from the Gly-Ser mutation in the highly charged RGE environment. Both CD and NMR real-time folding results indicate that peptide ERSEQ displays a comparatively slower folding rate than peptide DRSER, suggesting that the Gly-Ser mutation may lead to a larger interference in folding than the natural interruption in a similar RSE context. Our studies suggest that unlike the rigid GPO environment, the abundant R(K)GE(D) motif may provide a more flexible sequence environment that better accommodates mutations as well as interruptions, while the electrostatic interactions contribute to its stability. These results shed insight into the molecular features of the highly charged motif and may aid the design of collagen biomimetic peptides containing important biological sites.

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Section: Introductionmentioning

confidence: 99%

“…Meanwhile, peptide models of natural interruptions have also displayed localized alterations in conformation. 8,21,22 However, the molecular basis underlying the contradictory functional consequences of Gly mutations and natural interruptions is still poorly understood.…”

Section: Introductionmentioning

confidence: 99%

CD and NMR investigation of collagen peptides mimicking a pathological Gly-Ser mutation and a natural interruption in a similar highly charged sequence context

Sun

Liu

et al. 2015

Protein Science

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“…In the model, Phe222 forms a hydrophobic cluster with the corresponding residues in the other strands, as previously observed in other interruptions, where these inter-chain interactions stabilize the triple helix, and can favor the assembly of the trimer in the correct register. 15,17,20 Substitution of Cys would remove this stabilizing interaction, thus changing the structural and dynamical properties of the trimer. The mutation could also perturb protein-protein interactions (heat shock protein 47 binds at the mutation site).…”

Section: Case Reportmentioning

confidence: 99%

X-Linked Glomerulopathy Due to COL4A5 Founder Variant

Barua

Stella

et al. 2018

American Journal of Kidney Diseases

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Alport syndrome is a rare hereditary disorder caused by rare variants in 1 of 3 genes encoding for type IV collagen. Rare variants in COL4A5 on chromosome Xq22 cause X-linked Alport syndrome, which accounts for ∼80% of the cases. Alport syndrome has a variable clinical presentation, including progressive kidney failure, hearing loss, and ocular defects. Exome sequencing performed in 2 affected related males with an undefined X-linked glomerulopathy characterized by global and segmental glomerulosclerosis, mesangial hypercellularity, and vague basement membrane immune complex deposition revealed a COL4A5 sequence variant, a substitution of a thymine by a guanine at nucleotide 665 (c.T665G; rs281874761) of the coding DNA predicted to lead to a cysteine to phenylalanine substitution at amino acid 222, which was not seen in databases cataloguing natural human genetic variation, including dbSNP138, 1000 Genomes Project release version 01-11-2004, Exome Sequencing Project 21-06-2014, or ExAC 01-11-2014. Review of the literature identified 2 additional families with the same COL4A5 variant leading to similar atypical histopathologic features, suggesting a unique pathologic mechanism initiated by this specific rare variant. Homology modeling suggests that the substitution alters the structural and dynamic properties of the type IV collagen trimer. Genetic analysis comparing members of the 3 families indicated a distant relationship with a shared haplotype, implying a founder effect.

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“…Full-length collagen sequences contain multiple binding sites exhibiting variable integrin affinities 9,34,35 that effectively precludes an accurate energetic characterization. Synthetic collagen-like triple-helical peptides (THP) represent a suitable model for biophysical 5,7,30,[36][37][38][39] and functional studies 9,34,35 of collagen. In an effort to elucidate the forces that promote a1 I binding to collagen and the origins of E317A enhanced activity observed in adhesion assays toward collagens I and IV (Supporting Information Fig.…”

Section: Energetics Of Wild Type A1 I and E317a Interactions With A Cmentioning

confidence: 99%

Intrinsic local destabilization of the C‐terminus predisposes integrin α1 I domain to a conformational switch induced by collagen binding

et al. 2016

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Integrin-collagen interactions play a critical role in a myriad of cellular functions that include immune response, and cell development and differentiation, yet their mechanism of binding is poorly understood. There is increasing evidence that conformational flexibility assumes a central role in the molecular mechanisms of protein-protein interactions and here we employ NMR hydrogen-deuterium exchange (HDX) experiments to explore the impact of slower timescale dynamic events. To gain insight into the mechanisms underlying collagen-induced conformational switches, we have undertaken a comparative study between the wild type integrin a1 I and a gain-of-function E317A mutant. NMR HDX results suggest a relationship between regions exhibiting a reduced local stability in the unbound I domain and those that undergo significant conformational changes upon binding. Specifically, the aC and a7 helices within the C-terminus are at the center of such major perturbations and present reduced local stabilities in the unbound state relative to other structural elements. Complementary isothermal titration calorimetry experiments have been performed to derive complete thermodynamic binding profiles for association of the collagen-like triple-helical peptide with wild type a1 I and E317A mutant. The differential energetics observed for E317A are consistent with the HDX experiments and support a model in which intrinsically destabilized regions predispose Abbreviations: a1 I, alpha1 I domain; NMR, nuclear magnetic resonance; HDX, hydrogen-deuterium exchange; HSCQ, heteronuclear single-quantum correlation; MIDAS, metal ion-dependent adhesion site; THP, collagen-like triple helical peptide; ITC, isothermal titration calorimetry; CD, circular dichroism.Additional Supporting Information may be found in the online version of this article.Integrin-collagen interactions play a critical role in cell adhesion processes. To gain insight into the mechanisms underlying collageninduced conformational switches, we have undertaken a comparative NMR study between the human integrin a1 I and a gain-offunction E317A mutant. Our results, supported by thermodynamic measurements, suggest that intrinsically destabilized regions facilitate conformational rearrangements of integrin I domain. This study highlights the importance of exploring slow dynamics to delineate allosteric and binding events. Published by Wiley-Blackwell. V C 2016 The Protein Society conformational rearrangement in the integrin I domain. This study highlights the importance of exploring different timescales to delineate allosteric and binding events.

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NMR Studies Demonstrate a Unique AAB Composition and Chain Register for a Heterotrimeric Type IV Collagen Model Peptide Containing a Natural Interruption Site

Cited by 23 publications

References 46 publications

CD and NMR investigation of collagen peptides mimicking a pathological Gly-Ser mutation and a natural interruption in a similar highly charged sequence context

CD and NMR investigation of collagen peptides mimicking a pathological Gly-Ser mutation and a natural interruption in a similar highly charged sequence context

X-Linked Glomerulopathy Due to COL4A5 Founder Variant

Intrinsic local destabilization of the C‐terminus predisposes integrin α1 I domain to a conformational switch induced by collagen binding

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