Involvement of PYK 2 in Angiotensin II Signaling of Vascular Smooth Muscle Cells

PYK2, a recently identified Ca -sensitive tyrosine kinase, has been implicated in extracellular signal-regulated kinase (ERK) activation via several G protein–coupled receptors. We have reported that angiotensin II (Ang II) induces Ca-dependent transactivation of the epidermal growth factor receptor (EGFR) which serves as a scaffold for preactivated c-Src and downstream adaptors (Shc/Grb2), leading to ERK activation in cultured rat vascular smooth muscle cells (VSMC). Herein we demonstrate the involvement of PYK2 in this cascade. Ang II rapidly induced tyrosine phosphorylation of PYK2, whose effect was completely inhibited by an AT 1 receptor antagonist and an intracellular Ca chelator. A Ca ionophore also induced PYK2 tyrosine phosphorylation to a level comparable with that by Ang II, whereas phorbol ester–induced phosphorylation was less than that by Ang II. Moreover, PYK2 formed a complex coprecipitable with catalytically active c-Src after Ang II stimulation. Although a selective EGFR kinase inhibitor completely abolished Ang II–induced recruitment of Grb2 to EGFR and markedly attenuated Ang II–induced ERK activation, it had no effect on Ang II–induced PYK2 tyrosine phosphorylation or its association with c-Src and Grb2. These data suggest that the AT 1 receptor uses Ca -dependent PYK2 to activate c-Src, thereby leading to EGFR transactivation, which preponderantly recruits Grb2 in rat VSMC. (Hypertension. 1999;33[part II]:201-206.)

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