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Yeast ubiquitin ligase LC-MS/MS

ABSTRACT: Ubiquitin ligases (E3s) serves as a key regulator for ubiquitylation-mediated pathway. The identification of the corresponding relationship between E3 and its substrates is challenging but required for understanding the regulatory network of ubiquitylation. The low abundance of ubiquitinated conjugates and high redundancy of E3s-substrates regulation made screening pretty hard. Herein, we combined SILAC-based quantitative proteomics with two contrary genetic methods (overexpression and knockout) in theory for E3 (Hrt3, F-box subunit of SCF complex) substrates screening.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Yihao Wang

LAB HEAD: Ping Xu

PROVIDER: PXD015654 | Pride | 2019-12-06

REPOSITORIES: Pride

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Quantitative Proteomics Combined with Two Genetic Strategies for Screening Substrates of Ubiquitin Ligase Hrt3.

Lan Qiuyan Q Wang Yihao Y Sun Zhen Z Li Yanchang Y Zhang Cheng C Chang Lei L Gao Yue Y Wu Junzhu J Wang Fuqiang F Xu Ping P

Journal of proteome research 20191223 1

Ubiquitin ligases (E3s) serve as key regulators for the ubiquitylation-mediated pathway. The identification of the corresponding relationship between E3 and its substrates is challenging but required for understanding the regulatory network of ubiquitylation. The low abundance of ubiquitinated conjugates and high redundancy of E3 substrate regulation made the screening pretty hard. Herein, we combined SILAC-based quantitative proteomics with two contrary genetic methods (overexpression and knock ...[more]

PMID: 31789040

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