Phosphorylation-induced conformational changes and functional regulation of the muscle-atrophy-associated ubiquitin ligase Cbl-b
| Project/Area Number |
23770121
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
MAITA Ayako 徳島大学, 大学院・ヘルスバイオサイエンス研究部, 助教 (60415106)
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| Research Collaborator |
UEJI Tatsuya 徳島大学, 大学院・栄養生命科学教育部博士前期過程, 大学院生
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| Project Period (FY) |
2011 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Keywords | 分子認識及び相互作用 / 構造生物学 / 翻訳後修飾 |
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| Research Abstract |
To elucidate the structural basis for enhancement of Cbl-b’s ubiquitin ligase activity through the phosphorylation, we initially constructed a protein expression system for Cbl-b^<39-465>Y363E mutant that mimics the phosphorylated state of Cbl-b. We succeed in the development of a protein purification protocol in order to generate pure protein in sufficient quantities for X-ray crystallographic studies. We carried out crystal screening of a Cbl-b^<39-465>Y363E purified by this protocol. However, Huang’s group reported the crystal structure of the phosphorylated c-Cbl, a homolog of Cbl-b, in January 2012. Thus, we changed the originally planned research project for the study focused on the interaction between the Cbl-b^<39-341>(hereafter called "Cbl-b TKB") and the phosphorylated IRS-1. We succeeded in the crystal structure determination of the Cbl-b TKB in complex with the short phosphopeptide mimetic of phospholylated IRS-1. It revealed the details of the binding mode of Cbl-b TKB to the phosphopeptide.
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Report
(3 results)
Research Products
(6 results)
