Regulation of cell adhesion and type VII collagen binding by the beta3 chain short arm of laminin-5: effect of its proteolytic cleavage.
The basement membrane protein laminin-5 (Lm5), a heterotrimer of alpha3 (or alpha3A), beta3, and gamma2 chains, regulates cellular adhesion and motility. Here we examined the proteolysis and biological function of the laminin beta3 chain. First, we found that the beta3 chain of Lm5 is cleaved at its N-terminal, short arm by an endogenous proteinase(s) in normal human keratinocytes and some other cell lines. To examine the effect of beta3 chain cleavage, we expressed a wild-type Lm5 and two Lm5 mutants with partially deleted beta3 chains in HEK293 cells. Experiments with the purified Lm5 forms demonstrated that the deletion of the beta3 short arm or its N-terminal domain LN decreases the cell adhesion activity of Lm5, but does not significantly affect the motility activity. A recombinant beta3 short arm protein enhanced integrin-mediated cell adhesion to Lm5 by binding to an unidentified cell receptor. It was also found that the laminin EGF-like domain of the beta3 short arm is a binding site for type VII collagen. These results suggest that the beta3 short arm is involved not only in the matrix assembly of Lm5, but also in its cell adhesion activity. The proteolytic cleavage of the beta3 chain may modulate these functions of Lm5 in vivo.
Nakashima
Yukiko
Y
Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama 244-0813.
Kariya
Yoshinobu
Y
Yasuda
Chie
C
Miyazaki
Kaoru
K
D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002465:Cell Movement; D024143:Collagen Type VII / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000378:metabolism; D010447:Peptide Hydrolases; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D024143:Collagen Type VII; D016023:Integrins; D010447:Peptide Hydrolases; D011994:Recombinant Proteins; D000097794:Kalinin