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. 2012 May;166(2):457–475. doi: 10.1111/j.1476-5381.2011.01778.x

Table 2.

Summary of affinity methods – kinases

Protein target
Protein kinases Drug/inhibitor analogues Compound class Affinity method Separation method Detection method/MS Identified proteins Reference
CDKs (ser/thr-specific) purvalanol 2,6,9-tri-substituted purine PAC SDS-PAGE microsequencing 9 protein kinases (Knockaert et al., 2000)
CDKs/KSK-3 (ser/thr-specific) paullones PAC SDS-PAGE microsequencing GSK-3α, GSK-3β, MDHM (Knockaert et al., 2002)
CDK2 (ser/thr-specific) PHA-539136 cyclopropylpyrazole PAC SDS-PAGE/2D PAGE MALDI MS Cdk2, HSP-90α, 90β, 78, 70, 27, enolase, tubulin–β chain (Lolli et al., 2003)
CDK2 (ser/thr-specific) (R)-Roscovitine pegylated roscovitine PAC SDS-PAGE MALDI MS, Western blotting (Bach et al., 2005)
p38/MAPK family SB 203580 pyridinyl imidazole PAC 16-BAC/SDS-PAGE MALDI MS (Godl et al., 2003)
PKC family (ser/thr-specific) GF109203X (BisI) bisindolylmaleimide PAC 16-BAC/SDS-PAGE MALDI MS 17 protein kinases (Brehmer et al., 2004)
PKCα family BisIII bisindolylmaleimide PAC /SAC IAP, SDS-PAGE ESI IT MS/MS PKC-R, GSK3-β, CaMKII, adenosine kinase, CDK2 (Saxena et al., 2008) (Saxena et al., 2009)
Bcr-Abl (tyr-specific) PD180970/PP58 pyrido[2,3,-d]pyrimidine PAC 16-BAC/SDS-PAGE MALDI MS/ESI Q-TOF MS 35 protein kinases (Wissing et al., 2004)
Bcr-Abl (tyr-specific) imatinib, nilotinib, dasatinib, INNO-406 2-phenylamino pyrimidine PAC SDS-PAGE ESI Q-TOF MS multiple kinases (Rix et al., 2007) (Rix et al., 2011) (Li et al., 2010)
EGFR (tyr-specific) Gefitinib/AX14596 PAC 16-BAC/SDS-PAGE MALDI MS 20 protein kinases (Brehmer et al., 2005)
PDGFR, VEGFR2 (tyr-specific) SU6668 indolinone PAC 16-BAC/SDS-PAGE MALDI MS 8 protein kinases (Godl et al., 2005)
Jurkat, HCT-116, A549 cell kinomes multiple PAC SDS-PAGE ESI Q-TOF MS 140 protein kinases (Wissing et al., 2007)
HeLa S3 S- and M- phase kinomes multiple PAC SDS-PAGE, Mudpit SILAC, LTQ-Orbitrap MS 219 protein kinases (Daub et al., 2008)
MV4–11 leukaemia, HCT116, MDA-MB-435S kinomes VI16741/VI16743/VI16832 pyrido[2,3,-d]pyrimidine PAC SDS-PAGE SILAC, LTQ-Orbitrap MS 200 protein kinases (Oppermann et al., 2009)
Multiple mouse and human cell lines and primary tissues multiple PAC iTRAQ, LTQ-Orbitrap MS 307 protein kinases (Bantscheff et al., 2007)
CLL cells multiple PAC iTRAQ, LTQ-Orbitrap MS multiple kinases (Kruse et al., 2011)
HepG2 hepatocarcinoma, human placenta staurosporine azido, biotin SAC Streptavidin chromatography LTQ-Orbitrap MS 100 kinases (Fischer et al., 2010)

IAP. immuno affinity purification.