A Novel Binding Site in Collagen Type III for Integrins α1β1 and α2β1

• Published in: The Journal of biological chemistry 2005-09, Vol.280 (37), p.32512-32520
• Main Authors: Kim, Jiyeun Kate, Xu, Yi, Xu, Xuejun, Keene, Douglas R., Gurusiddappa, Sivashankarappa, Liang, Xiaowen, Wary, Kishore K., Höök, Magnus
• Format: Article
• Language: English
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M502431200

Previously identified high affinity integrin-binding motifs in collagens, GFOGER and GLOGER, are not present in type III collagen. Here, we first characterized the binding of recombinant I domains from integrins α1 and α2 (α1I and α2I) to fibrillar collagen types I-III and showed that each I domain bound to the three types of collagens with similar affinities. Using rotary shadowing followed by electron microscopy, we identified a high affinity binding region in human type III collagen recognized by α1I and α2I. Examination of the region revealed the presence of two sequences that contain the critical GER motif, GROGER and GAOGER. Collagen-like peptides containing these two motifs were synthesized, and their triple helical nature was confirmed by circular dichroism spectroscopy. Experiments show that the GROGER-containing peptide was able to bind both α1I and α2I with high affinity and effectively inhibit the binding of α1I and α2I to type III and I collagens, whereas the GAOGER-containing peptide was considerably less effective. Furthermore, the GROGER-containing peptide supported adhesion of human lung fibroblast cells when coated on a culture dish. Thus, we have identified a novel high affinity binding sequence for the collagen-binding integrin I domains.