Cell-associated activation of latent transforming growth factor-beta by calpain.
Transforming growth factor-beta (TGF-beta) is normally secreted in a latent form, and plasmin-mediated proteolytic cleavage of latency-associated peptide (LAP), a component of latent TGF-beta complex that makes the complex inactive, activates latent TGF-beta. In the present study, we investigated the possible involvement of calpain, one of the cysteine proteases, in the activation of latent TGF-beta. When recombinant latent TGF-beta was incubated with calpain (1-10 u/ml) in a test tube, calpain cleaved LAP and released mature TGF-beta from the latent complex. When calpain was applied to cultured bovine capillary endothelial (BCE) cells, a low concentration of calpain (0.05-0.1 u/ml) inhibited the migration and proliferation of the cells, and these inhibitory effects were abrogated by anti-TGF-beta antibody as well as by calpain inhibitor peptide, but not by alpha2-antiplasmin, a specific inhibitor of plasmin. Active TGF-beta was detected in the conditioned medium of BCE cells collected in the presence of calpain. Chemical cross-linking of (125)I-calpain to BCE cells followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicated that calpain bound to the cell surface through chondroitinase ABC-sensitive proteoglycan. In addition, treatment of the BCE cells with chondroitinase ABC abrogated the inhibitory effect of calpain on the migration of these cells. Our data thus suggest that calpain is able to activate latent TGF-beta through a mechanism independent of plasmin. This activation is efficient in the presence of cells, and calpain binds to the cell surface via proteoglycan and activates latent TGF-beta, which is targeted to the same surface.
Abe
M
M
Department of Vascular Biology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Japan.
Oda
N
N
Sato
Y
Y
D000818:Animals; D002154:Calpain / Q000494:pharmacology*; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002810:Chondroitinases and Chondroitin Lyases / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D002154:Calpain; D002810:Chondroitinases and Chondroitin Lyases