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Am J Physiol Renal Physiol 264: F532-F539, 1993;
0363-6127/93 $5.00
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AJP - Renal Physiology, Vol 264, Issue 3 532-F539, Copyright © 1993 by American Physiological Society

ARTICLES

Two-chain urokinase, receptor, and type 1 inhibitor in cultured human mesangial cells

W. F. Glass 2nd, J. I. Kreisberg and D. A. Troyer
Audie L. Murphy Memorial Veterans Hospital, Department of Pathology, University of Texas Health Science Center, San Antonio 78284.

Urokinase-type plasminogen activator (u-PA), its receptor (u-PAR), and type 1 inhibitor (PAI-1) in cultured human mesangial cells were investigated. Treatment with phospholipase C (PLC) released plasminogen activators [with relative mol wt (M(r)) of 55,000 and 100,000] and u-PAR into the culture medium. By Western blot, both u-PA and PAI-1 were present in the M(r) 100,000 band. Since PAI-1 binds only active, two-chain u-PA (tcu-PA), formation of the M(r) 100,000 band reflects conversion of the single-chain, proenzyme form of u-PA (scu-PA) to tcu-PA. Immunofluorescence staining of whole cells demonstrated the presence of PAI-1, u-PA, and u-PAR. Immunofluorescence staining and Western blot analysis showed enrichment of PAI-1, u-PA, and u-PAR in a preparation of substratum-attached extracellular matrix and membrane proteins termed adhesion plaques. Using a chromogenic assay, we found that PAI-1 expression in adhesion plaques exceeded that of u-PA. We conclude that cultured human mesangial cells produce receptor-bound u-PA/PAI-1 complexes localized to adhesion plaques.


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[Abstract] [Full Text] [PDF]


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