A method for determining ubiquitin chain length in cells

2016 Fiscal Year Final Research Report

• Project/Area Number: 15H06882
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Functional biochemistry
• Research Institution: Tokyo Metropolitan Institute of Medical Science
• 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)
• Project Period (FY): 2015-08-28 – 2017-03-31
• Keywords: ユビキチン

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.

Free Research Field

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