2007 Fiscal Year Final Research Report Summary
Biochemical characterization of small GTP binding protein in the brain of insect
| Project/Area Number |
18580055
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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 |
Applied entomology
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| Research Institution | Kobe University |
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Principal Investigator |
UNO Tomohide Kobe University, Agrobioscience, Associate professor (80240852)
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| Project Period (FY) |
2006 – 2007
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| Keywords | insect / brain / small GTP binding protein / antibody / Bombyx mori / rab |
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| Research Abstract |
Small GTPases of the Rab proteins are key regulators of membrane trafficking. It was reported that rab 8 from Bombyx mori (BRab8) was phosphorylated by protein kinase C (PKC), which were expressed in mammalian cells and partially purified (Uno, et. al., 2006). To determine the phosphorylated amino acid residues of BRab8, seven serine and four threonin residues were shown as candidates to be phosphorylated. So, six deleted or mutated fragments containing cDNAs for BRab8 were constructed using PCR. The obtained fragments were inserted into expression vectors. Fusion-proteins were expressed in E.coli and partially purified using affinity chromatographies. These proteins were phosphorylated by PKC and gamma ^<32>P-ATP in vitro, electrophoresed and analyzed by a bioimaging analyzer. As a result, one serine-132 of BRab8 was specifically phosphorylated. Further, polyclonal antibody was isolated by immunizing with rab8 protein. From immunoblotting and staining phosphorylated proteins, Rab8 was found to be phosphorylated in vivo.
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[Journal Article] Bioconversion by functional P450 .1A9 and IClof Anguillus japonica2007
Author(s)
Uno, T, Okamoto, S, Masuda, S, Imaisihi, H, Nakamura, M, Kanamaru, K, Yamagata, H, El-Kady, WA, Kaminishi Y, Itakura, T
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Journal Title
Comprehensive Biochemistry and Physiology C 147
Pages: 278-285
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Rab GTPase, small GTP binding proteins, from the brain of Bombyx mon2008
Author(s)
Keisuke, Hata, Le, Thi Dieu Trang, Tsubasa, Moriwaki, Takuya, Nakada, Susumu, Hiragaki, Michihiro, Takagi, Kengo, Kanamaru, Hiroshi, Yamagata, Makio, Takeda, Tomohide Uno
Organizer
Asia-pacific congress of Sericulture and Insect Biotechnology 2008 (APSERI 2008)
Place of Presentation
Nagoya University
Year and Date
2008-03-22
Description
「研究成果報告書概要(欧文)」より
