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PMID: 21700206 Published · ppublish English Journal Article

In situ kinase profiling reveals functionally relevant properties of native kinases.

Chemistry & biology ·Vol. 18 ·No. 6 ·2011-06-24 ·Pages 699-710

Patricelli MP, Nomanbhoy TK, Wu J, Brown H, Zhou D, Zhang J, Jagannathan S, Aban A, Okerberg E, Herring C, Nordin B, Weissig H, Yang Q, Lee JD, Gray NS, Kozarich JW

Abstract

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.

MeSH Terms
Adenosine Triphosphate/chemistry Cell Line, Tumor Dasatinib Humans MAP Kinase Kinase 5/antagonists & inhibitors,metabolism Protein Binding Protein Kinase Inhibitors/chemistry,pharmacology Protein Kinases/chemistry,genetics,metabolism Pyrimidines/chemistry,pharmacology Recombinant Proteins/antagonists & inhibitors,genetics,metabolism Thiazoles/chemistry,pharmacology raf Kinases/antagonists & inhibitors,genetics,metabolism
Chemicals
Protein Kinase Inhibitors Pyrimidines Recombinant Proteins Thiazoles Adenosine Triphosphate Protein Kinases raf Kinases MAP Kinase Kinase 5 MAP2K5 protein, human Dasatinib
Authors & Affiliations
16 authors, click to expand affiliations / ORCID
Patricelli Matthew P
ActivX Biosciences, La Jolla, CA 92037, USA.
Nomanbhoy Tyzoon K
Wu Jiangyue
Brown Heidi
Zhou David
Zhang Jianming
Jagannathan Subadhra
Aban Arwin
Okerberg Eric
Herring Chris
Nordin Brian
Weissig Helge
Yang Qingkai
Lee Jiing-Dwan
Gray Nathanael S
Kozarich John W
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Article Info
Journal
Chemistry & biology
Abbr.
Chem Biol
ISSN
1879-1301
DOI
10.1016/j.chembiol.2011.04.011
Published
2011-06-24
Pages
699-710
Language
English
Region
United States
NLM ID
9500160
PMCID
PMC3142620
Subset
IM
Grants
NCI NIH HHS · R01 CA130876 · United States
NCI NIH HHS · R01 CA130876-03 · United States
NIGMS NIH HHS · P41 GM079575-02 · United States
NCI NIH HHS · R01 CA130876-04 · United States
NIGMS NIH HHS · P41 GM079575-01 · United States
NIGMS NIH HHS · P41 GM079575 · United States
NCI NIH HHS · R01 CA130876-02 · United States
NCI NIH HHS · R01 CA130876-01A1 · United States
NIGMS NIH HHS · P41 GM079575-03S1 · United States
NIGMS NIH HHS · P41 GM079575-03 · United States
Corrections
CommentIn
PMID: 21700204
External Links
PubMed source DOI: 10.1016/j.chembiol.2011.04.011 PMC Full Text
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