Identification of integrin-dependent and -independent cell adhesion domains in COOH-terminal globular region of laminin-5 alpha 3 chain.
Laminin-5 is an isoform of laminin that consists of alpha 3, beta 3, and gamma 2 chains and has potent cell adhesion- and cell migration-promoting activities. In this study, five subdomains in the COOH-terminal globular (G) domain of human laminin alpha 3 chain were individually expressed in Escherichia coli, and their biological activities were investigated. Recombinant G2, G4, and G5 domains promoted adhesion to plastic plates of HT1080 fibrosarcoma cells, A431 epidermoid carcinoma cells, and ECV304 vascular endothelial cells. For the cell adhesion activity, the G2 domain required a divalent cation and heat-sensitive conformation more strongly than G4 and G5. The cell adhesion to G2 but not G4 and G5 was effectively inhibited by an anti-integrin alpha 3 antibody. A cell adhesion sequence of 22 amino acids, alpha 3G2A, that was homologous to the integrin alpha 3 beta 1-binding sequence GD-6 of laminin alpha 1 chain was identified within the G2 structure. The cell adhesion to alpha 3G2A peptide was also inhibited by the anti-integrin alpha 3 antibody. The cell adhesion to G2, alpha 3G2A, G4, and G5 was strongly inhibited by heparin, but that to native laminin-5 was inhibited less effectively. Moreover, G5 potently stimulated chemotactic migration of rat liver epithelial cells in Boyden chambers, but G2 and G4 did not. These results indicate that the G domain of laminin alpha 3 contains multiple cell binding sites with different mechanisms and different functions. The G2 domain seems to recognize integrin alpha 3 beta 1, whereas G4 and G5 may interact with heparin-like molecules on cell surface.
Mizushima
H
H
Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, Japan.
Takamura
H
H
Miyagi
Y
Y
Kikkawa
Y
Y
Yamanaka
N
N
Yasumitsu
H
H
Misugi
K
K
Miyazaki
K
K
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D002465:Cell Movement / Q000187:drug effects; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D016023:Integrins / Q000502:physiology*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D051381:Rats; D011915:Rats, Inbred BUF; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D006493:Heparin; D016023:Integrins; D011994:Recombinant Proteins; D000097794:Kalinin