A method for determining ubiquitin chain length in cells
| Project/Area Number |
15H06882
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| Research Category |
Grant-in-Aid for Research Activity Start-up
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional biochemistry
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| Research Institution | Tokyo Metropolitan Institute of Medical Science |
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Principal Investigator |
TSUCHIYA Hikaru 公益財団法人東京都医学総合研究所, 生体分子先端研究分野, 研究員 (90760132)
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| Project Period (FY) |
2015-08-28 – 2017-03-31
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| Project Status |
Completed (Fiscal Year 2016)
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| Budget Amount *help |
¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | ユビキチン / タンパク質分解 |
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| Outline of Final Research Achievements |
We describe a novel versatile method for assessing chain length of substrate-attached polyubiquitin chains. We named this method, which combines a high-affinity probe for ubiquitin and partial trypsin digestion of ubiquitin chains, ‘ubiquitin chain protection from trypsinization (Ub-ProT)’. Using this method,we analyzed the ubiquitin chain architecture of ligand-activated epidermal growth factor receptor (EGFR) in human cells. By combining mass spectrometry-based ubiquitin chain quantification, deubiquitinase-based analysis, and Ub-ProT, we revealed that EGFR is rapidly modified by K63-linked tetra- to hexa-ubiquitin chains following EGF treatment.
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Report
(3 results)
Research Products
(4 results)
