Identification of novel substrates of protein kinase ROCK
| Project/Area Number |
12680693
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | RIKEN (2001)
Kyoto University (2000) |
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Principal Investigator |
MAEKAWA Midori RIKEN Center for Developmental Biology, Cell adhesion and tissue patterning group, Research Specialist, 高次構造形成研究グループ, 専門職研究員 (70314185)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Rho / ROCK / Y-27632 |
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| Research Abstract |
The aim of this project is to clarify the signal pathways downstream of Rho and Rho-dependent serine/threonine protein kinase ROCK, through the screening and characterization of novel substrates of ROCK.
I used two-dimensional gel electrophoresis and autoradiography to detect ROCK targets whose phosphorylation is Induced by LPA and inhibited by a specific inhibitor of ROCK, Y-27632 in three LPA-sensitive cell lines. In the NIE115 neuroblastoma cells, I detected a 200-KDa protein. The protein was identified as a -fodrin through the analysis of MALDl-TOF mass spectrometry. Further investigation, however, showed that the LPA-sensitivity of the phosphorylation of α-fodrin was very Iow. In the Swiss3T3 fibroblasf cells, I defecfed a 25-KDa profein. This protein was suggested to be myosin light chain (MLC) from the result of MALDI-TOF mass spectrometry, and further western blotting analysis confirmed this result. MLC has been reported as a target downstream of ROCK. In summary, novel substrates of ROCKhad not been identiffed in this project. However, since the validity of this approach was ascertained, further application of this method to various other cells that respond to LPA in different ways, will be useful to identify the targets of ROCK.
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Report
(3 results)
Research Products
(9 results)
