A Hexylchloride-Based Catch-and-Release System for Chemical Proteomic Applications

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• 作者：Jennifer L. Brigham ; B. Gayani K. Perera ; Dustin J. Maly
• 刊名：ACS Chemical Biology
• 出版年：2013
• 出版时间：April 19, 2013
• 年：2013
• 卷：8
• 期：4
• 页码：691-699
• 全文大小：427K
• 年卷期：v.8,no.4(April 19, 2013)
• ISSN：1554-8937

文摘

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.