| Project/Area Number |
13558084
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Functional biochemistry
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
HAMAKUBO Takao RESEARCH CENTER FOR ADVANCED SCIENCE AND TECNOLOGY, PROFESSOR, 先端科学技術研究センター, 教授 (90198797)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Toshiya GRELAN PHARMACEUTICAL CO. LTD., RESEARCH ASSOCIATE, 研究開発部開発研究所, 研究員
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2002: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 2001: ¥8,900,000 (Direct Cost: ¥8,900,000)
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| Keywords | MEMBRANE PROTEIN / BACULOVIRUS / NUCLEAR RECEPTOR / MONOCLONAL ANTIBODY / GATA / GPCR / コレステロール代謝 / 蛋白質相互作用 |
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| Research Abstract |
Membrane proteins are the major targets for the genome based drug discovery in the post-genomic era. In this research we developed a novel expression system for membrane proteins using baculovirus and utilized it to raise specific antibodies against membrane proteins and nuclear receptor superfamily. We found that a novel property of baculovirus expression system that some membrane proteins can be displayed on the budded virus itself. To date we have observed the membrane display of several ER proteins such as sterol regulatory element-binding proteirt-2 (SREBP-2), SREBP cleavage-activating protein (SCAP), HMG-CoA reductase, presenilin, and nicastrin. All of them had proper activities. Also we have succeeded to reconstitute the trimeric G proteins and G protein coupled receptors on the budded baculovirus. Those reconstituted GPCR had high affinity for ligand binding. Thus this viral expression system considered to be very useful for the analysis of membrane proteins. We then utilized this system for making antibody against membrane proteins. We have been succeeded to generate specific monoclonal antibodies against several membrane proteins such as transporter, SCAP, and nicastrin. On the other hand, using gp64 fusion protein, a membrane protein of baculovirus itself, series of monoclonal antibodies has been generated against superfamily of nuclear receptor. We have been achieved to raise 30 monoclonal antibodies out of 48 nuclear receptor family proteins using this system. Thus we have been developed novel viral display system for membrane proteins and its application for antibody generation.
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