| Autor: |
Kunhong Xiao, Jinpeng Sun, Jihee Kim, Rajagopal, Sudarshan, Bo Zhai, Villén, Judit, Haas, Wilhelm, Kovacs, Jeffrey J., Shukla, Arun K., Hara, Makoto R., Hernandez, Marylens, Lachmann, Alexander, Zhao, Shan, Lin, Yuan, Yishan Cheng, Mizuno, Kensaku, Ma!ayan, Avi, Gygi, Steven P., Lefkowitz, Robert J. |
| Předmět: |
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| Zdroj: |
Proceedings of the National Academy of Sciences of the United States of America; 8/24/2010, Vol. 107 Issue 34, p15299-15304, 6p |
| Abstrakt: |
βArrestin-mediated signaling downstream of seven transmembrane receptors (7TMR5) is a relatively new paradigm for signaling by these receptors. We examined changes in protein phosphorylation occurring when HEK293 cells expressing the angiotensin II type 1A receptor (AT1aR) were stimulated with the β-arrestin-biased ligand Sar1, lIe4, 11e8-angiotensin (Sll), a ligand previously found to signal through β-arrestin-dependent, G protein-independent mechanisms. Using a phospho-antibody array containing 46 antibodies against signaling molecules, we found that phosphorylation of 35 proteins increased upon Sll stimulation. These 511-mediated phosphorylation events were abrogated after depletion of β-arrestin 2 through siRNA-mediated knockdown. We also performed an MS-based quantitative phosphoproteome analysis after SlI stimulation using a strategy of stable isotope labeling of amino acids in cell culture (SILAC). We identified 1,555 phosphoproteins (4,552 unique phosphopeptides), of which 171 proteins (222 phosphopeptides) showed increased phosphorylation, and 53 (66 phosphopeptides) showed decreased phosphorylation upon Sil stimulation of the AT1aR. This study identified 38 protein kinases and three phosphatases whose phosphorylation status changed upon 511 treatment. Using computational approaches, we performed systembased analyses examining the β-arrestin--mediated phosphoproteome including construction of a kinase-substrate network for β-arrestinmediated AT1aR signaling. Our analysis demonstrates that β-arrestindependent signaling processes are more diverse than previously appreciated. Notably, our analysis identifies an AT1 aR-mediated cytoskeletal reorganization network whereby β-arrestin regulates phosphorylation of several key proteins, including cot ilin and slingshot This study provides a system-based view of β-arrestin-mediated phosphorylation events downstream of a 7TMR and opens avenues for research in a rapidly evolving area of 7TMR signaling. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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