期刊
ACS CHEMICAL BIOLOGY
卷 13, 期 8, 页码 1932-1937出版社
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.8b00178
关键词
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资金
- Marie Curie Career Integration Grant of the Radboud University in Nijmegen
- Institute of Molecules and Materials (IMM) of the Radboud University in Nijmegen
Mapping proteins at a specific subcellular location is essential to gaining detailed insight on local protein dynamics. We have developed an enzymatic strategy to label proteins on a subcellular level using arylamine N-acetyltransferase (NAT). The NAT enzyme activates an arylhydroxamic acid functionality into a nitrenium ion that reacts fast, covalently, and under neutral conditions with nucleophilic residues of neighboring proteins. The electron density on the aromatic ring proved important for probe activation as strong labeling was only observed with an arylhydroxamic acid bearing an electron donating substituent. We further demonstrate that, using this electron rich arylhydroxamic acid, clear labeling was achieved on a subcellular level in living cells that were transfected with a genetically targeted NAT to the nucleus or the cytosol.
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