In Situ Kinase Profiling Reveals Functionally Relevant Properties of Native Kinases

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• 作者：Matthew  ; P. Patricelli¹ ; ^{mattp@activx.com"" rel=""nofollow} ; Tyzoon  ; K. Nomanbhoy¹ ; Jiangyue Wu¹ ; Heidi Brown¹ ; David Zhou¹ ; Jianming Zhang² ; Subadhra Jagannathan¹ ; Arwin Aban¹ ; Eric Okerberg¹ ; Chris Herring¹ ; Brian Nordin¹ ; Helge Weissig¹ ; Qingkai Yang³ ; Jiing-Dwan Lee³ ; Nathanael  ; S. Gray² ; John  ; W. Kozarich¹
• 刊名：Chemistry & Biology
• 出版年：2011
• 出版时间：24 June 2011
• 年：2011
• 卷：18
• 期：6
• 页码：699-710
• 全文大小：530 K

文摘

Summary

Protein kinases are intensely studied mediators of cellular signaling, yet important questions remain regarding their regulation and in vivo properties. Here, we use a probe-based chemoprotemics platform to profile several well studied kinase inhibitors against >200 kinases in native cell proteomes and reveal biological targets for some of these inhibitors. Several striking differences were identified between native and recombinant kinase inhibitory profiles, in particular, for the Raf kinases. The native kinase binding profiles presented here closely mirror the cellular activity of these inhibitors, even when the inhibition profiles differ dramatically from recombinant assay results. Additionally, Raf activation events could be detected on live cell treatment with inhibitors. These studies highlight the complexities of protein kinase behavior in the cellular context and demonstrate that profiling with only recombinant/purified enzymes can be misleading.