Sensitive multiplexed analysis of kinase activities and activity-based kinase identification Export

@article{citeulike:5900707, abstract = {Constitutive activation of one or more kinase signaling pathways is a hallmark of many cancers. Here we extend the previously described mass spectrometry–based KAYAK approach by monitoring kinase activities from multiple signaling pathways simultaneously. This improved single-reaction strategy, which quantifies the phosphorylation of 90 synthetic peptides in a single mass spectrometry run, is compatible with nanogram to microgram amounts of cell lysate. Furthermore, the approach enhances kinase monospecificity through substrate competition effects, faithfully reporting the signatures of many signaling pathways after mitogen stimulation or of basal pathway activation differences across a panel of well-studied cancer cell lines. Hierarchical clustering of activities from related experiments groups peptides phosphorylated by similar kinases together and, when combined with pathway alteration using pharmacological inhibitors, distinguishes underlying differences in potency, off-target effects and genetic backgrounds. Finally, we introduce a strategy to identify the kinase, and even associated protein complex members, responsible for phosphorylation events of interest.}, author = {Kubota, Kazuishi and Anjum, Rana and Yu, Yonghao and Kunz, Ryan C. and Andersen, Jannik N. and Kraus, Manfred and Keilhack, Heike and Nagashima, Kumiko and Krauss, Stefan and Paweletz, Cloud and Hendrickson, Ronald C. and Feldman, Adam S. and Wu, Chin-Lee and Rush, John and Villen, Judit and Gygi, Steven P.}, citeulike-article-id = {5900707}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nbt.1566}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nbt.1566}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/19801977}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=19801977}, day = {04}, doi = {10.1038/nbt.1566}, issn = {1087-0156}, journal = {Nature Biotechnology}, keywords = {good, kinase, method, ms}, month = {October}, number = {10}, pages = {933--940}, posted-at = {2009-11-12 09:22:32}, priority = {2}, publisher = {Nature Publishing Group}, title = {Sensitive multiplexed analysis of kinase activities and activity-based kinase identification}, url = {http://dx.doi.org/10.1038/nbt.1566}, volume = {27}, year = {2009} }

TY - JOUR ID - citeulike:5900707 L3 - citeulike-article-id:5900707 N2 - Constitutive activation of one or more kinase signaling pathways is a hallmark of many cancers. Here we extend the previously described mass spectrometry–based KAYAK approach by monitoring kinase activities from multiple signaling pathways simultaneously. This improved single-reaction strategy, which quantifies the phosphorylation of 90 synthetic peptides in a single mass spectrometry run, is compatible with nanogram to microgram amounts of cell lysate. Furthermore, the approach enhances kinase monospecificity through substrate competition effects, faithfully reporting the signatures of many signaling pathways after mitogen stimulation or of basal pathway activation differences across a panel of well-studied cancer cell lines. Hierarchical clustering of activities from related experiments groups peptides phosphorylated by similar kinases together and, when combined with pathway alteration using pharmacological inhibitors, distinguishes underlying differences in potency, off-target effects and genetic backgrounds. Finally, we introduce a strategy to identify the kinase, and even associated protein complex members, responsible for phosphorylation events of interest. IS - 10 JF - Nature Biotechnology SN - 1087-0156 EP - 940 TI - Sensitive multiplexed analysis of kinase activities and activity-based kinase identification PB - Nature Publishing Group VL - 27 SP - 933 KW - good KW - kinase KW - method KW - ms AU - Kubota, Kazuishi AU - Anjum, Rana AU - Yu, Yonghao AU - Kunz, Ryan AU - Andersen, Jannik AU - Kraus, Manfred AU - Keilhack, Heike AU - Nagashima, Kumiko AU - Krauss, Stefan AU - Paweletz, Cloud AU - Hendrickson, Ronald AU - Feldman, Adam AU - Wu, Chin-Lee AU - Rush, John AU - Villen, Judit AU - Gygi, Steven PY - 2009/10/04/ UR - http://dx.doi.org/10.1038/nbt.1566 ER -