Tyrosine phosphorylation of the dense plaque protein paxillin is regulated during smooth muscle contraction. Wang, Zhonglin, Fredrick. M. Pavalko, and Susan J. Gunst. Dept. of Physiology and Biophysics, Indiana Univ. School of Medicine, Indianapolis, IN, 46202
APStracts 3:0164C, 1996.
Regulation of the attachment of actin filaments to the cell membrane at membrane-associated dense plaque (MADP) sites could allow smooth muscle cells to modulate their cytostructure in response to changes in external stress. In this study, changes in the tyrosine phosphorylation of the MADP protein, paxillin, were measured by Western blot during the contraction and relaxation of tracheal smooth muscle strips. Tyrosine phosphorylation of paxillin increased by 3-4 fold with a time course similar to force development during contractile stimulation with ACh, 5-hydroxytryptamine and KCl, and decreased during washout of contractile stimuli and during relaxation induced by forskolin. Immunoprecipitation of muscle extracts with multiple rounds of anti-phosphotyrosine antibody removed approximately 20% of the total paxillin in resting muscles, and about 60% of paxillin in muscles maximally stimulated with ACh. These results provide the first evidence associating the tyrosine phosphorylation of paxillin with the active contraction of smooth muscle, or with any functional response of a fully differentiated tissue in vivo. The results are consistent with a role for MADP proteins in the regulation of force development in smooth muscle.

Received 29 January 1996; accepted in final form 8 May 1996.
APS Manuscript Number C58-6.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 28 May 96