Mixed peptide–chitosan membranes to mimic the biological activities of a multifunctional laminin α1 chain LG4 module

Abstract

Laminin α1 chain LG4 module is multifunctional and interacts with syndecans and integrin α2β1 via AG73 (RKRLQVQLSIRT) and EF-I (DYATLQLQEGRLHFMFDLG) sites, respectively. Here, we conjugated the AG73 and EF1zz (ATLQLQEGRLHFXFDLGKGR, X: Nle) peptides on a chitosan membrane in various ratios to develop an LG4 mimic biomaterial. The AG73–chitosan membrane promoted strong cell attachment with membrane ruffling and the EF1zz–chitosan membrane promoted integrin-mediated cell adhesion with well-organized actin stress fibers. When AG73 and EF1zz were conjugated on a chitosan membrane with 1:9 molar ratio, the mixed peptide–chitosan membrane promoted the strong cell attachment and neurite outgrowth similar to that on the recombinant LG4 protein. Well-organized actin stress fibers and vinculin accumulated focal contacts were observed in the cells attached on the AG73:EF1zz (molar ratio = 1:9)–chitosan membrane. These results suggest that the mixed peptide–chitosan membrane interacts with both syndecans and integrin α2β1 and mimics the cell adhesion of a multifunctional LG4 protein. The mixed peptide–chitosan approach has potential as a multifunctional biomaterial for cell and tissue engineering.