title: In situ kinase profiling reveals functionally relevant properties of native kinases.
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.
authors.affiliation: ActivX Biosciences, La Jolla, CA 92037, USA.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Patricelli
authors.forename: Tyzoon K
authors.initials: TK
authors.lastname: Nomanbhoy
authors.forename: Jiangyue
authors.initials: J
authors.lastname: Wu
authors.forename: Heidi
authors.initials: H
authors.lastname: Brown
authors.forename: David
authors.initials: D
authors.lastname: Zhou
authors.forename: Jianming
authors.initials: J
authors.lastname: Zhang
authors.forename: Subadhra
authors.initials: S
authors.lastname: Jagannathan
authors.forename: Arwin
authors.initials: A
authors.lastname: Aban
authors.forename: Eric
authors.initials: E
authors.lastname: Okerberg
authors.forename: Chris
authors.initials: C
authors.lastname: Herring
authors.forename: Brian
authors.initials: B
authors.lastname: Nordin
authors.forename: Helge
authors.initials: H
authors.lastname: Weissig
authors.forename: Qingkai
authors.initials: Q
authors.lastname: Yang
authors.forename: Jiing-Dwan
authors.initials: JD
authors.lastname: Lee
authors.forename: Nathanael S
authors.initials: NS
authors.lastname: Gray
authors.forename: John W
authors.initials: JW
authors.lastname: Kozarich
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D045744:Cell Line, Tumor; D000069439:Dasatinib; D006801:Humans; D048671:MAP Kinase Kinase 5 / Q000037:antagonists & inhibitors / Q000378:metabolism; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles / Q000737:chemistry / Q000494:pharmacology; D048490:raf Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D011994:Recombinant Proteins; D013844:Thiazoles; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D048490:raf Kinases; D048671:MAP Kinase Kinase 5; C482563:MAP2K5 protein, human; D000069439:Dasatinib