Identification of biologically active sequences in the laminin alpha2 chain G domain.
Laminin alpha2 chain is specifically expressed in the basement membrane surrounding muscle and nerve. We screened biologically active sequences in the mouse laminin alpha2 chain G domain using 110 soluble peptides by the peptide-coated plate and the peptide-conjugated Sepharose bead assays. Fourteen peptides showed cell attachment activity in either or both assays. Cell attachment to A2G94 (YFDGTGFAKAVG) was inhibited by anti-integrin beta1 antibody, suggesting that the peptide promotes an integrin beta1-mediated cell attachment. Five peptides promoted PC12 cell neurite outgrowth. Since A2G10 (SYWYRIEASRTG) promoted strong cell attachment in the bead assay but showed slight activity in the plate assay, we conjugated A2G10 to chitosan membranes which increase cell attachment activity of the peptides via conformational stability. A2G10-chitosan membrane promoted an integrin alpha6beta1-mediated cell attachment and spreading with well-organized actin stress fibers and neurite outgrowth. These active peptides are useful for evaluating the molecular mechanisms of laminin-receptor interactions.
Urushibata
Shunsuke
S
Laboratory of Clinical Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192-0392, Japan.
Hozumi
Kentaro
K
Ishikawa
Masaya
M
Katagiri
Fumihiko
F
Kikkawa
Yamato
Y
Nomizu
Motoyoshi
M
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000378:metabolism; D007797:Laminin / Q000737:chemistry* / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D016501:Neurites / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D016716:PC12 Cells; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D051381:Rats
D019012:Integrin beta1; D007797:Laminin; D009842:Oligopeptides