Immunoaffinity profiling of tyrosine phosphorylation in cancer cells.

Article Details


Rush J, Moritz A, Lee KA, Guo A, Goss VL, Spek EJ, Zhang H, Zha XM, Polakiewicz RD, Comb MJ

Immunoaffinity profiling of tyrosine phosphorylation in cancer cells.

Nat Biotechnol. 2005 Jan;23(1):94-101. Epub 2004 Dec 12.

PubMed ID
15592455 [ View in PubMed

Tyrosine kinases play a prominent role in human cancer, yet the oncogenic signaling pathways driving cell proliferation and survival have been difficult to identify, in part because of the complexity of the pathways and in part because of low cellular levels of tyrosine phosphorylation. In general, global phosphoproteomic approaches reveal small numbers of peptides containing phosphotyrosine. We have developed a strategy that emphasizes the phosphotyrosine component of the phosphoproteome and identifies large numbers of tyrosine phosphorylation sites. Peptides containing phosphotyrosine are isolated directly from protease-digested cellular protein extracts with a phosphotyrosine-specific antibody and are identified by tandem mass spectrometry. Applying this approach to several cell systems, including cancer cell lines, shows it can be used to identify activated protein kinases and their phosphorylated substrates without prior knowledge of the signaling networks that are activated, a first step in profiling normal and oncogenic signaling networks.

DrugBank Data that Cites this Article

NameUniProt ID
L-lactate dehydrogenase B chainP07195Details
Pyruvate kinase PKMP14618Details
Tubulin beta chainP07437Details
T-cell surface glycoprotein CD3 epsilon chainP07766Details
ALK tyrosine kinase receptorQ9UM73Details
L-lactate dehydrogenase A chainP00338Details
Low-density lipoprotein receptorP01130Details
Glyceraldehyde-3-phosphate dehydrogenaseP04406Details
Tyrosine-protein phosphatase non-receptor type 1P18031Details
Glutathione reductase, mitochondrialP00390Details
Cysteine--tRNA ligase, cytoplasmicP49589Details
Inosine-5'-monophosphate dehydrogenase 2P12268Details
Activated CDC42 kinase 1Q07912Details
Malate dehydrogenase, mitochondrialP40926Details
Tyrosine-protein kinase LckP06239Details
Trifunctional purine biosynthetic protein adenosine-3P22102Details
Phosphatidylinositol 3-kinase regulatory subunit alphaP27986Details
Spermine synthaseP52788Details
Insulin receptor substrate 1P35568Details
Protein deglycase DJ-1Q99497Details
Mitogen-activated protein kinase 14Q16539Details
Tyrosine-protein phosphatase non-receptor type 11Q06124Details
Microtubule-associated protein 1AP78559Details
Tyrosine-protein kinase ZAP-70P43403Details
Heat shock protein HSP 90-betaP08238Details
Thioredoxin reductase 1, cytoplasmicQ16881Details
Fatty acid-binding protein, epidermalQ01469Details
Cyclin-dependent kinase 6Q00534Details
Succinyl-CoA ligase [ADP-forming] subunit beta, mitochondrialQ9P2R7Details
Nicotinamide phosphoribosyltransferaseP43490Details
Wiskott-Aldrich syndrome proteinP42768Details
Far upstream element-binding protein 1Q96AE4Details
T-cell surface glycoprotein CD3 zeta chainP20963Details
Signal transducer and activator of transcription 3P40763Details
Tyrosine-protein kinase JAK3P52333Details
60S ribosomal protein L23P62829Details
60S ribosomal protein L8P62917Details
Proteasome subunit alpha type-2P25787Details
Proteasome subunit alpha type-3P25788Details
Proteasome subunit beta type-4P28070Details
Dual specificity tyrosine-phosphorylation-regulated kinase 1AQ13627Details
Actin-related protein 3P61158Details
Tyrosine-protein kinase LynP07948Details
Phosphoglycerate kinase 1P00558Details
Ubiquitin-like modifier-activating enzyme 1P22314Details
Histone acetyltransferase KAT2AQ92830Details
Hepatocyte growth factor-regulated tyrosine kinase substrateO14964Details
Transferrin receptor protein 1P02786Details
60 kDa heat shock protein, mitochondrialP10809Details
Heterogeneous nuclear ribonucleoprotein HP31943Details
Heterogeneous nuclear ribonucleoprotein H3P31942Details
Receptor of activated protein C kinase 1P63244Details
Stress-induced-phosphoprotein 1P31948Details
Eukaryotic translation initiation factor 6P56537Details
Heterogeneous nuclear ribonucleoprotein FP52597Details
Heat shock 70 kDa protein 4P34932Details
Splicing factor 3A subunit 3Q12874Details
40S ribosomal protein S13P62277Details
Probable ATP-dependent RNA helicase DDX5P17844Details
Serine/threonine-protein kinase PRP4 homologQ13523Details
ATP-citrate synthaseP53396Details
Thioredoxin reductase 1, cytoplasmicQ9JMH6Details