Molecular underpinnings of integrin binding to collagen-mimetic peptides containing vascular Ehlers-Danlos syndrome-associated substitutions.
Collagens carry out critical extracellular matrix (ECM) functions by interacting with numerous cell receptors and ECM components. Single glycine substitutions in collagen III, which predominates in vascular walls, result in vascular Ehlers-Danlos syndrome (vEDS), leading to arterial, uterine, and intestinal rupture and an average life expectancy of <50 years. Collagen interactions with integrin α2β1 are vital for platelet adhesion and activation; however, how these interactions are impacted by vEDS-associated mutations and by specific amino acid substitutions is unclear. Here, we designed collagen-mimetic peptides (CMPs) with previously reported Gly → Xaa (Xaa = Ala, Arg, or Val) vEDS substitutions within a high-affinity integrin α2β1-binding motif, GROGER. We used these peptides to investigate, at atomic-level resolution, how these amino acid substitutions affect the collagen III-integrin α2β1 interaction. Using a multitiered approach combining biological adhesion assays, CD, NMR, and molecular dynamics (MD) simulations, we found that these substitutions differentially impede human mesenchymal stem cell spreading and integrin α2-inserted (α2I) domain binding to the CMPs and were associated with triple-helix destabilization. Although an Ala substitution locally destabilized hydrogen bonding and enhanced mobility, it did not significantly reduce the CMP-integrin interactions. MD simulations suggested that bulkier Gly → Xaa substitutions differentially disrupt the CMP-α2I interaction. The Gly → Arg substitution destabilized CMP-α2I side-chain interactions, and the Gly → Val change broke the essential Mg2+ coordination. The relationship between the loss of functional binding and the type of vEDS substitution provides a foundation for developing potential therapies for managing collagen disorders.
['Hoop|Cody L|CL|0000-0003-4441-8321', 'Kemraj|Allysa P|AP|', 'Wang|Baifan|B|', 'Gahlawat|Sonal|S|', 'Godesky|Madison|M|', 'Zhu|Jie|J|', 'Warren|Haley R|HR|', 'Case|David A|DA|', 'Shreiber|David I|DI|', 'Baum|Jean|J|']
D019943:Amino Acid Substitution D001665:Binding Sites D002448:Cell Adhesion D002460:Cell Line D003094:Collagen D004535:Ehlers-Danlos Syndrome D006801:Humans D038982:Integrin alpha2beta1 D059630:Mesenchymal Stem Cells D062105:Molecular Docking Simulation D010455:Peptides D011485:Protein Binding
integrin collagen vascular Ehlers-Danlos syndrome cell spreading cell surface receptor collagen mutations connective tissue disorder extracellular matrix protein protein-protein interaction triple helix