Wang, Jacky. Relation of conformational and dynamic differences in collagen sequences to alpha2I domain integrin binding affinity. Retrieved from https://doi.org/doi:10.7282/T37D2X0B
DescriptionThe α2I domain of integrin has been identified to bind to the recognition motifs in collagen, GFOGER and GAOGER, with the former having a higher binding affinity than the latter. Here we employ nuclear magnetic resonance spectroscopy and circular dichroism spectroscopy to study the structural differences between the GFOGER and GAOGER collagen model peptides (CMP) triple helices. As a consequence of the limitations from using synthetic CMPs, we have also used recombinant collagen proteins expressed from E. coli as a way to efficiently screen mutations in CMPs and provide a platform to probe binding affinities using enzyme-linked immunosorbent assay. Results from this work show that the residues in GAOGER and GFOGER triple helices have few differences in dynamics, however dynamics between strands in a collagen triple helix may play a role in modulation of binding affinity. Attempts at more efficient analysis of varying CMPs using recombinant proteins have yielded us fusion proteins that can be used with enzyme-linked immunoabsorbent assay and produce recombinant CMPs.