Angiotensin II stimulation of rapid protein tyrosine phosphorylation and protein kinase activation in rat aortic smooth muscle cells.
In cultured rat aortic smooth muscle cells, angiotensin II (AII) treatment led to increased tyrosine phosphorylation of cellular proteins with apparent molecular masses of 42, 44, 75, and 120 kDa, respectively, as assessed by antiphosphotyrosine immunoblotting. Increased protein tyrosine phosphorylation was observed within 1 min of AII addition and was maximal by 30 min. The overall pattern of AII-stimulated protein tyrosine phosphorylation was distinct from that observed following treatment of rat aortic smooth muscle cells with platelet-derived growth factor-BB. Specific antibodies were used to identify the AII-stimulated 42- and 44-kDa tyrosine-phosphorylated proteins as the "mitogen-activated protein kinases," p42mapk and p44mapk, respectively. Raf-1, a 70-74-kDa serine/threonine protein kinase, was not tyrosine-phosphorylated in response to AII but was found to be hyperphosphorylated as evidenced by retarded protein mobility in SDS gel analysis. Taken together, these data indicate that AII binding to vascular smooth muscle cells leads to rapid activation of a complex cascade of protein kinases, including protein kinase C, Raf-1, MAP kinases, and an undefined intracellular protein tyrosine kinase(s) that may be coordinately involved in signal transduction leading to cell proliferation.
Department of Cardiovascular Biochemistry, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543.
C J
CJ
Molloy
D S
DS
Taylor
H
H
Weber
D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D001011:Aorta; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002455:Cell Division; D002478:Cells, Cultured; D004789:Enzyme Activation; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000201:enzymology*; D010766:Phosphorylation; D011233:Precipitin Tests; D011494:Protein Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D019908:Proto-Oncogene Proteins c-raf; D051381:Rats; D017207:Rats, Sprague-Dawley; D014443:Tyrosine / Q000378:metabolism*
D011518:Proto-Oncogene Proteins; D000804:Angiotensin II; D014443:Tyrosine; D011494:Protein Kinases; D019908:Proto-Oncogene Proteins c-raf; D017871:Calcium-Calmodulin-Dependent Protein Kinases