Monitoring Native p38伪:MK2/3 Complexes via Trans Delivery of an ATP Acyl Phosphate Probe

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• 作者：Eric S. Okerberg ; Heidi E. Brown ; Lauro Minimo ; Senait Alemayehu ; Jonathan Rosenblum ; Matt Patricelli ; Tyzoon Nomanbhoy ; John W. Kozarich
• 刊名：Journal of the American Chemical Society
• 出版年：2014
• 出版时间：March 26, 2014
• 年：2014
• 卷：136
• 期：12
• 页码：4664-4669
• 全文大小：324K
• 年卷期：v.136,no.12(March 26, 2014)
• ISSN：1520-5126

文摘

Here we describe a chemical proteomics strategy using ATP acyl phosphates to measure the formation of a protein:protein complex between p38伪 and mapkap kinases 2 and/or 3. Formation of the protein:protein complex results in a new probe labeling site on p38伪 that can be used to quantify the extent of interaction in cell lysates and the equilibrium binding constant for the interaction in vitro. We demonstrate through RNA interference that the labeling site is dependent on formation of the protein:protein complex in cells. Further, we identify that active-site-directed, small-molecule inhibitors of MK2/3 selectively inhibit the heterodimer-dependent probe labeling, whereas p38伪 inhibitors do not. These findings afford a new method to evaluate p38伪 and MK2/3 inhibitors within native biological systems and a new tool for improved understanding of p38伪 signaling pathways.