Mixed peptide-chitosan membranes to mimic the biological activities of a multifunctional laminin alpha1 chain LG4 module.
Laminin alpha1 chain LG4 module is multifunctional and interacts with syndecans and integrin alpha2beta1 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 alpha2beta1 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.
Hozumi
Kentaro
K
Laboratory of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan.
Yamagata
Natsumi
N
Otagiri
Dai
D
Fujimori
Chikara
C
Kikkawa
Yamato
Y
Kadoya
Yuichi
Y
Nomizu
Motoyoshi
M
D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D048271:Chitosan / Q000494:pharmacology*; D004492:Edetic Acid / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000737:chemistry* / Q000494:pharmacology*; D008567:Membranes, Artificial; D051379:Mice; D018716:Molecular Mimicry / Q000187:drug effects*; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D016716:PC12 Cells; D010446:Peptide Fragments / Q000494:pharmacology; D010455:Peptides / Q000494:pharmacology*; D051381:Rats; D011994:Recombinant Proteins / Q000494:pharmacology; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism
C105786:AG 73; D000199:Actins; D007797:Laminin; D008567:Membranes, Artificial; D010446:Peptide Fragments; D010455:Peptides; D011994:Recombinant Proteins; D016596:Vinculin; C084098:laminin A; D006493:Heparin; D048271:Chitosan; D004492:Edetic Acid