Method for finding E2-E3 specifically mediating target protein ubiquitination reaction based on known E1
A target protein, specific technology, applied in the field of biochemistry and molecular biology, can solve the problems of terror and unclear ubiquitin modification mechanism
Inactive Publication Date: 2013-02-20
JIANGSU UNIV
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Abstract
The invention relates to the fields of biochemistry and molecular biology, and specifically relates to a method forfinding an ubiquitin ligase E3 specifically identifying a target protein and a E2/E3 system for regulating target protein ubiquitination reaction from a number of E3 of human cells. According to the invention, an in-vitro ubiquitination analysis system is established; a known ubiquitin activating enzyme E1 and HelaS100 fraction are adopted as E3 sources, and a ligase E2 mediating the target protein ubiquitination reaction is screened; and a multiple chromatographic column combination is combined with mass spectrometric analysis, such that the ubiquitin ligase E3 specifically identifying the target protein and the E2/E3 system regulating target protein ubiquitination modification are searched from human cells. With the method, the target-protein-identifying substrate recognition factor which is ubiquitin ligase E3 can be conveniently and accurately found, and a E2/E3 system mediating target protein ubiquitination modification can be established.
Application Domain
Microbiological testing/measurementBiological testing
Technology Topic
Chromatographic columnUbiquitin +9
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