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ACS Chem Biol. 2013 Apr 19;8(4):691-9. doi: 10.1021/cb300623a. Epub 2013 Jan 23.

A hexylchloride-based catch-and-release system for chemical proteomic applications.

Author information

1
Department of Chemistry, University of Washington, Seattle, WA 98195, USA.

Abstract

Bioorthogonal ligation methods that allow the selective conjugation of fluorophores or biotin to proteins and small molecule probes that contain inert chemical handles are an important component of many chemical proteomic strategies. Here, we present a new catch-and-release enrichment strategy that utilizes a hexylchloride group as a bioorthogonal chemical handle. Proteins and small molecules that contain a hexylchloride tag can be efficiently captured by an immobilized version of the self-labeling protein HaloTag. Furthermore, by using a HaloTag fusion protein that contains a protease cleavage site, captured proteins can be selectively eluted under mild conditions. We demonstrate the utility of the hexylchloride-based catch-and-release strategy by enriching protein kinases that are covalently and noncovalently bound to ATP-binding site-directed probes from mammalian cell lysates. Our catch-and-release system creates new possibilities for profiling enzyme families and for the identification of the cellular targets of bioactive small molecules.

PMID:
23305300
PMCID:
PMC3631427
DOI:
10.1021/cb300623a
[Indexed for MEDLINE]
Free PMC Article

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