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Chemical Protein Ubiquitylation with Preservation of the Native Cysteine Residues
Author(s) -
Yang Kun,
Li Guorui,
Gong Ping,
Gui Weijun,
Yuan Libo,
Zhuang Zhihao
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201600042
Subject(s) - ubiquitin , cysteine , proliferating cell nuclear antigen , native chemical ligation , biochemistry , chemistry , chemical biology , deubiquitinating enzyme , microbiology and biotechnology , biology , dna , enzyme , gene
We report a cysteine‐based ligation strategy for generating a monoubiquitylated protein while preserving the native cysteine residues on the acceptor protein. In monoubiquitylation of proliferating cell nuclear antigen (PCNA) this method circumvents the need to mutate the native cysteine residues on PCNA. The chemically ubiquitylated PCNA contains a noncleavable linkage of the same length as the native isopeptide linkage. It also retains the normal function of the native Ub‐PCNA in stimulating the ATPase activity of replication factor C (RFC) and lesion bypass synthesis by Polη. This method may be adapted for chemical ubiquitylation of other proteins and for site‐specific modification of a target protein at a specific site through sulfhydryl chemistry.