| Project/Area Number |
13480215
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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 | 一般 |
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| Research Field |
Biophysics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SUTOH Kazuo Professor, Department of Life Sciences,The University of Tokyo, 大学院・総合文化研究科, 教授 (20111453)
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| Co-Investigator(Kenkyū-buntansha) |
KON Takahide Research Assistant, Department of Life Sciences, The University of Tokyo, 大学院・総合文化研究科, 助手 (30332620)
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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 |
¥14,900,000 (Direct Cost: ¥14,900,000)
Fiscal Year 2002: ¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 2001: ¥7,600,000 (Direct Cost: ¥7,600,000)
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| Keywords | Molecular Motor / Processivity / Human myosin / Unconventional Myosin / Recombinant Protein / ミオシン / 滑り運動 / プロセッシブ運動 / モーティリティーアッセイ |
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| Research Abstract |
Motor proteins belong one of the classes according to their structure and behavior ; two-headed non-processive, two-headed processive, one-headed non-processive and one-headed processive. Myosin II is in the first class. Myosin V and VI belong to the second class. Myosin I belongs to the third class. One of the kinesin superfamily proteins belong to the last class. In human myosins, myosin IIIA, myosin IIIB, myosin IXA, and myosin IXA are supposed to belong to the last class, although it has not yet been determined experimentally if these myosins are actually one-headed and processive. We have cloned full-length cDNA of human myosin IIIA, IIIB, and IXB from the human brain cDNA library. Cloned myosin IIIA and IXB cDNAs show that there are several types of alternative splicing. We have expressed these cDNAs including several splice variants in Sf9 or high five cells. From the transfected cells, we have purified the expressed proteins for further biochemical and biophysical analyzes. At this stage, we are only partially successful to isolate the proteins due to the formation of inclusion bodies. Various purification procedures have been tried to increase the solubility of expressed proteins. Further studies are required to finally identify the motor characteristics of myosin IIIA, IIIB and IXB.
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