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Vol. 304, Issue 1, 63-70, January 2003

Calpain Mediates Progressive Plasma Membrane Permeability and Proteolysis of Cytoskeleton-Associated Paxillin, Talin, and Vinculin during Renal Cell Death

Xiuli Liu and Rick G. Schnellmann

Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas (X.L.); and Department of Pharmaceutical Sciences, Medical University of South Carolina, Charleston, South Carolina (R.G.S.)

The goal of the present study was to determine the role of calpain in changes in plasma membrane permeability and cytoskeleton-associated paxillin, vinculin, talin, and alpha -actinin levels during acute renal cell death. The mitochondrial inhibitor antimycin A or hypoxia produced graded plasma membrane permeability in renal proximal tubules (RPTs), first allowing propidium iodide (PI, molecular mass 668 Da) influx and then lactate dehydrogenase (LDH, molecular mass 130 kDa) release. Cytoskeleton-associated paxillin levels decreased concomitantly with PI influx and before LDH release, whereas cytoskeleton-associated talin and vinculin levels decreased concomitantly with LDH release. Cytoskeleton-associated alpha -actinin levels did not change during antimycin A exposure or hypoxia. Purified µ-calpain cleaved paxillin, talin, vinculin, but not alpha -actinin. The dissimilar calpain inhibitors 3-(4-iodophenyl)-2-mercapto-(Z)-2-propenoic acid (PD150606) or chloroacetic acid N'-[6,7-dichloro-4-phenyl)-3-oxo-3,4-dihydroquinoxalin-2-yl] hydrazide (SJA7029) preserved cytoskeleton-associated paxillin, talin, and vinculin levels and prevented PI influx and LDH release in antimycin A-exposed or hypoxic RPTs. These results suggest that calpain mediates increased plasma membrane permeability and hydrolysis of cytoskeleton-associated paxillin, vinculin, and talin during renal cell death.

0022-3565/03/3041-0063$07.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2003 by The American Society for Pharmacology and Experimental Therapeutics


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