2000 Fiscal Year Final Research Report Summary
PROTEOMIC ANALYSIS OF SYNAPTIC MOLECULAR STRUCTURES AND FUNCTIONAL REGULATION BY PROTEIN PHOSPHORYLATION
| Project/Area Number |
11680768
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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 |
Neurochemistry/Neuropharmacology
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| Research Institution | RIKEN (2000)
Fujita Health University (1999) |
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Principal Investigator |
TANIGUCHI Hisaaki RIKEN HARIMA INSTITUTE, TEAMLEADER, 翻訳後修飾による動的調節機構研究チーム, チームリーダー(研究職) (10257636)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUBARA Mamoru RIKEN HARIMA INSTITUTE, RESEARCHER, 翻訳後修飾による動的調節機構研究チーム, 連携研究員 (90288481)
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| Project Period (FY) |
1999 – 2000
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| Keywords | MASS SPECTROMETRY / PROTEOMICS / SYNAPSE / POSTTRANSLATIONAL MODIFICATIONS PROTEIN PHOSPHORYLATION / SIGNAL TRANSDUCTION / シグナル伝達 |
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| Research Abstract |
The present research project has two aims. (1)To elucidate the regulatory mechanisms of presynaptic neurotransmitter release by protein phosphorylation. (2)To analyze the synaptic molecular structure including sinal transduction pathways. For the former, ultra-sensitive mass spectrometric analysis should be applied to analyze the protein phosphorylation of proteins in the synaptic vesicles and in the SNARE complex. For the latter, the same mass spectrometric method is applied to analyze the component proteins found in the growth cones as well as the postsynaptic densities. At the same time, the protein phosphorylation of these proteins found in the latter study should be analyzed. To achieve these studies, we have established a proteomics facility centered on five mass spectrometers. This facility includes also various automatic robots such as gel picking robot and in-gel digestion robot. We have established a high-throughput analysis system consisting of various robots. A triple-stage quadrupole mass spectrometer was combined with a capillary HPLC to conduct phosphopeptide analysis based on precursor scanning method. During these research we could identify more than 50 phosphorylation sites in a brain-specific microtubule-associated protein MAP1B.We have also identified several syntaxin-binding proteins.
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Research Products
(14 results)
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[Publications] Uedaira, H., Morii, H., Ishimura, M., Taniguchi, H., Namba, K., and Vonderviszt, F: "Domain organization of flagellar hook protein from Salmonella typhimurium"FEBS Lett. 421. 203-207 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Takasaki, A., Hayashi, N., Matsubara, M., Yamauchi, E., and Taniguchi, H: "Identification of the calmodulin-binding domain of neuron-specific protein kinase C substrate protein CAP-23/NAP-22 : Direct involvement of protein myristoylation in calmodulin-target protein interaction"J.Biol, Chem. 274. 11848-11853 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Manenti, S., Yamauchi, E., Sorokine, O., Knibiehler, M., Van Dorsselaer, A., Taniguchi, H., Ducommun, B., and Darbon, J.-M.: "Phosphorylation of the myristoylated PKC substrate MARCKS by the cyclin E-Cdk2 complex in vitro"Biochem. J.. 340(Pt3). 775-782 (1999)
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「研究成果報告書概要(欧文)」より
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[Publications] Morimatsu, M., Nakamura, A., Sumiyoshi, H., Sakaba, N., Taniguchi, H., Kohama, K., and Higashi-Fujime, S.: "The molecular structure of the fastest myosin from green algae, Chara"Biochem Biophys Res Commun. 270. 147-152 (2000)
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「研究成果報告書概要(欧文)」より
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[Publications] Yamauchi, E.Muramoto, K., Kuroda, Y.and Taniguchi, H.: "Site-specific phosphorylation and dephosphorylation of microtubule-associated protein MAP1B during neuronal development in Neural development(Uemura, K.Kawamura, K.and Yazaki, T., eds.)"Springer-Verlag, Tokyo. 421-424 (1999)
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「研究成果報告書概要(欧文)」より
