© 1995 American Heart Association, Inc.
Articles |
Angiotensin II Activates pp60c-src in Vascular Smooth Muscle Cells
From the Division of Cardiology (M.I., T.I., B.C.B.), Department of Medicine, University of Washington, Seattle, and the Department of Pathology (M.B.M., B.S., K.E.B.), Emory University, Atlanta, Ga.
Correspondence to Dr Bradford C. Berk, Division of Cardiology, 357710, University of Washington, Seattle, WA 98195. E-mail bcberk@u.washington.edu.
Abstract The angiotensin II type-1
(AT1) receptor, a G protein–coupled receptor, lacks
intrinsic kinase activity. However, recent data show that
angiotensin II (Ang II) stimulates tyrosine
phosphorylation of phospholipase C-
1 (PLC-1),
Stat91 (one of the signal transducers and activators of
transcription), and paxillin in vascular smooth muscle cells. The
tyrosine kinases responsible for these phosphorylation
events are unknown. Src family kinases have been shown to
phosphorylate PLC-1 and to be activated by G
protein–coupled receptors. We hypothesized that
pp60c-src associates with the
AT1 receptor and is activated after Ang II
stimulation of smooth muscle cells. We immunoprecipitated
pp60c-src from Ang II–stimulated
vascular smooth muscle cells and measured
pp60c-src activity by
autophosphorylation and by
phosphorylation of enolase. Both assays demonstrated an
approximately threefold increase in
pp60c-src activity within 1 minute.
A similar increase in Ang II–stimulated
pp60c-src activity was observed in
Chinese hamster ovary cells transfected with the AT1
receptor but not in untransfected cells. These data are the first to
show that pp60c-src is
activated by Ang II. To determine if
pp60c-src associated with the
AT1 receptor, the AT1 receptor was
immunoprecipitated (with two different antibodies), and Western blots
were performed with two different
anti-pp60c-src antibodies. No
pp60c-src was detected. In
addition, direct interaction between the AT1 receptor and
pp60c-src could not be demonstrated
by using a glutathione S-transferase (GST)-AT1 fusion
protein to bind proteins from cell lysates stimulated by Ang II. In
combination with recent findings that
anti-pp60c-src antibodies
inhibit Ang II–mediated PLC-1 phosphorylation, our
data suggest an important role for
pp60c-src in Ang II signal
transduction.
Key Words: angiotensin II • signal transduction • vascular smooth muscle • Src kinase
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K. K. Griendling, M. Ushio-Fukai, B. Lassegue, and R. W. Alexander Angiotensin II Signaling in Vascular Smooth Muscle: New Concepts Hypertension, January 1, 1997; 29(1): 366 - 370. [Abstract] [Full Text] [PDF]
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Y. Zou, I. Komuro, T. Yamazaki, R. Aikawa, S. Kudoh, I. Shiojima, Y. Hiroi, T. Mizuno, and Y. Yazaki Protein Kinase C, but Not Tyrosine Kinases or Ras, Plays a Critical Role in Angiotensin II-induced Activation of Raf-1 Kinase and Extracellular Signal-regulated Protein Kinases in Cardiac Myocytes J. Biol. Chem., December 27, 1996; 271(52): 33592 - 33597. [Abstract] [Full Text] [PDF]
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D.-F. Liao, J. L. Duff, G. Daum, S. L. Pelech, and B. C. Berk Angiotensin II Stimulates MAP Kinase Kinase Kinase Activity in Vascular Smooth Muscle Cells: Role of Raf Circ. Res., November 1, 1996; 79(5): 1007 - 1014. [Abstract] [Full Text]
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L. M. Luttrell, B. E. Hawes, T. van Biesen, D. K. Luttrell, T. J. Lansing, and R. J. Lefkowitz Role of c-Src Tyrosine Kinase in G Protein-coupled Receptorand Gbeta gamma Subunit-mediated Activation of Mitogen-activated Protein Kinases J. Biol. Chem., August 9, 1996; 271(32): 19443 - 19450. [Abstract] [Full Text] [PDF]
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B. Schieffer, W. G. Paxton, Q. Chai, M. B. Marrero, and K. E. Bernstein Angiotensin II Controls p21[IMAGE] Activity via pp60[IMAGE] J. Biol. Chem., April 26, 1996; 271(17): 10329 - 10333. [Abstract] [Full Text] [PDF]
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B. Schieffer, W. G. Paxton, M. B. Marrero, and K. E. Bernstein Importance of Tyrosine Phosphorylation in Angiotensin II Type 1 Receptor Signaling Hypertension, March 1, 1996; 27(3): 476 - 480. [Abstract] [Full Text]
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L. Guillemot, A. Levy, Z. J. Zhao, G. Bereziat, and B. Rothhut The Protein-tyrosine Phosphatase SHP-2 Is Required during Angiotensin II-mediated Activation of Cyclin D1 Promoter in CHO-AT1A Cells J. Biol. Chem., August 18, 2000; 275(34): 26349 - 26358. [Abstract] [Full Text] [PDF]
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K. C. Chung, J. Y. Sung, W. Ahn, H. Rhim, T. H. Oh, M. G. Lee, and Y. S. Ahn Intracellular Calcium Mobilization Induces Immediate Early Gene pip92 via Src and Mitogen-activated Protein Kinase in Immortalized Hippocampal Cells J. Biol. Chem., January 12, 2001; 276(3): 2132 - 2138. [Abstract] [Full Text] [PDF]
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P. P. Sayeski, M. S. Ali, S. J. Frank, and K. E. Bernstein The Angiotensin II-dependent Nuclear Translocation of Stat1 Is Mediated by the Jak2 Protein Motif 231YRFRR J. Biol. Chem., March 23, 2001; 276(13): 10556 - 10563. [Abstract] [Full Text] [PDF]
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A. Bonacchi, P. Romagnani, R. G. Romanelli, E. Efsen, F. Annunziato, L. Lasagni, M. Francalanci, M. Serio, G. Laffi, M. Pinzani, et al. Signal Transduction by the Chemokine Receptor CXCR3. ACTIVATION OF Ras/ERK, Src, AND PHOSPHATIDYLINOSITOL 3-KINASE/Akt CONTROLS CELL MIGRATION AND PROLIFERATION IN HUMAN VASCULAR PERICYTES J. Biol. Chem., March 23, 2001; 276(13): 9945 - 9954. [Abstract] [Full Text] [PDF]
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P. Rocic and P. A. Lucchesi Down-regulation by Antisense Oligonucleotides Establishes a Role for the Proline-rich Tyrosine Kinase PYK2 in Angiotensin II-induced Signaling in Vascular Smooth Muscle J. Biol. Chem., June 8, 2001; 276(24): 21902 - 21906. [Abstract] [Full Text] [PDF]
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