THE MECHANISM FOR THE ACTIVATION OF LATENT TGF-BETA DURING COCULTURE OF ENDOTHELIAL-CELLS AND SMOOTH-MUSCLE CELLS - CELL-TYPE-SPECIFIC TARGETING OF LATENT TGF-BETA TO SMOOTH-MUSCLE CELLS

Transforming growth factor-beta (TGF-beta) is secreted in a latent for m and activated during co-culture of endothelial cells and smooth musc le cells. Plasmin located on the surface of endothelial cells is requi red for the activation of latent TGF-beta (LTGF-beta) during co-cultur e, and the targeting of LTGF-beta to the cellular surface is requisite for its activation. In the present study, the cellular targeting of L TGF-beta was examined. We detected the specific binding of I-125-large LTGF-beta1 isolated from human platelets to smooth muscle cells but n ot to endothelial cells. A mAb against the latency-associated peptide (LAP) of large LTGF-beta1 complex, which blocked the binding of I-125- large LTGF-beta1 to smooth muscle cells, inhibited the activation of L TGF-beta during co-culture. The binding of I-125-large LTGF-beta1 coul d not be competed either by mannose-6-phosphate (300 muM) or by the sy nthetic peptide Arg-Gly-Asp-Ser (300 mug/ml). These results indicate t hat the targeting of LTGF-beta to smooth muscle cells is required for the activation of LTGF-beta during co-culture of endothelial cells and smooth muscle cells. The targeting of LTGF-beta to smooth muscle cell s is mediated by LAP, and the domain of LAP responsible for the target ing to smooth muscle cells may not be related to mannose-6-phosphate o r an Arg-Gly-Asp sequence, both of which have been previously proposed as candidates for the cellular binding domains within LAP.