Project/Area Number |
11680776
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Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Neurochemistry/Neuropharmacology
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Research Institution | Institute for Developmental Research, Aichi Human Service Center |
Principal Investigator |
T.SUZUKI Shintaro Institute for Developmental Research, Aichi Human Service Center, Department of Developmental Biology, director, 発生学部門, 部長 (20128432)
|
Co-Investigator(Kenkyū-buntansha) |
HIRANO Shinji Institute for Developmental Research, Aichi Human Service Center, Department of Development Developmental Biology, investigator, 発生学部門, 研究員 (80222248)
|
Project Period (FY) |
1999 – 2000
|
Project Status |
Completed (Fiscal Year 2000)
|
Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥2,000,000 (Direct Cost: ¥2,000,000)
|
Keywords | affinity chromatography / mass spectroscopy / protocadherin / binding proteins / synapse / immunofluorescence microscopy / electrophoresis / yeast two hybrid method / 生理機能 / 免疫沈降法 / 免疫蛍光染色法 / 電気泳動 / 蛍光染色 / モノクローナル抗体 / カドヘリン |
Research Abstract |
Identification and characterization of proteins that associate with the cytoplasmic domains of several protocadherins have been carried out. Candidate binding proteins were isolated by affinity chromatography using GST-fusion proteins of the cytoplasmic domains of protocadherins and were identified by mass-spectroscopy. Yeast two-hybrid method was also used to isolate the binding proteins. The results revealed candidate proteins mostly belonged to cytoskeletal proteins. Immuno-fluorescence microscopy and immunoprecipitation were, then, performed to show that the candidate proteins actually associated with protocadherins in vivo. Preliminary results showed promissing in several cases. However, a major obstacle is that good antibodies against the candidates are not available in most cases. We are now producing the antibodies against the candidates' to facilitate the experiments. We are also planning ectopic expression experiments of the candidates to clarify the biological role of protocadherins.
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