Cell-adhesive activity and receptor-binding specificity of the laminin-derived YIGSR sequence grafted onto Staphylococcal protein A.
Laminin contains multiple oligopeptide motifs to promote cell adhesion and migration. One of these motifs is YIGSR within the B1 chain. We reconstituted the cell-adhesive activity of YIGSR motif by grafting it onto a truncated form of the Staphylococcal protein A (designated tSPA) via cassette mutagenesis. When coated on a polystyrene surface, the YIGSR-grafted tSPA (YIGSR-tSPA) promoted attachment and spreading of mouse melanoma and human rhabdomyosarcoma cells, but not of hamster fibroblasts. The cell-adhesive activity of YIGSR-tSPA was abolished by amino acid substitution or scrambling of the inserted YIGSR sequence. Divalent cations Mn2+ and Mg2+, but not Ca2+, promoted the cell adhesion to YIGSR-tSPA. Interestingly, the YIGSR-tSPA-mediated cell adhesion was barely inhibited by the linear peptide CDPGYIGSR-NH2, but was strongly inhibited by the cyclic peptide CDPGYIGSRC and another peptide PEILDVPST, which is a specific inhibitor for integrin alpha 4 beta 1. Among various anti-integrin antibodies, anti-alpha 4 and anti-beta 1 antibodies specifically inhibited the cell adhesion to YIGSR-tSPA. In support of these observations, adhesion of rhabdomyosarcoma cells to intact laminin was also partially inhibited by synthetic PEILDVPST peptide and anti-alpha 4 antibody. These results, taken together, indicate that the YIGSR motif exerts its cell-adhesive activity through interaction with integrin alpha 4 beta 1.
Maeda
T
T
Research Institute, Osaka Medical Center for Maternal and Child Health.
Titani
K
K
Sekiguchi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002413:Cations, Divalent / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects*; D002460:Cell Line; D006224:Cricetinae; D006801:Humans; D039041:Integrin alpha4beta1; D016023:Integrins / Q000276:immunology / Q000502:physiology; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009842:Oligopeptides / Q000378:metabolism* / Q000494:pharmacology*; D015202:Protein Engineering; D012208:Rhabdomyosarcoma / Q000378:metabolism; D013205:Staphylococcal Protein A; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D039041:Integrin alpha4beta1; D016023:Integrins; D007797:Laminin; D009842:Oligopeptides; D013205:Staphylococcal Protein A; C081571:tyrosyl-isoleucyl-glycyl-seryl-arginine