Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics

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Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics. / Pan, Cuiping; Olsen, Jesper V; Daub, Henrik; Mann, Matthias.

In: Molecular and Cellular Proteomics, Vol. 8, No. 12, 2009, p. 2796-808.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Pan, C, Olsen, JV, Daub, H & Mann, M 2009, 'Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics', Molecular and Cellular Proteomics, vol. 8, no. 12, pp. 2796-808. https://doi.org/10.1074/mcp.M900285-MCP200

APA

Pan, C., Olsen, J. V., Daub, H., & Mann, M. (2009). Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics. Molecular and Cellular Proteomics, 8(12), 2796-808. https://doi.org/10.1074/mcp.M900285-MCP200

Vancouver

Pan C, Olsen JV, Daub H, Mann M. Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics. Molecular and Cellular Proteomics. 2009;8(12):2796-808. https://doi.org/10.1074/mcp.M900285-MCP200

Author

Pan, Cuiping ; Olsen, Jesper V ; Daub, Henrik ; Mann, Matthias. / Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics. In: Molecular and Cellular Proteomics. 2009 ; Vol. 8, No. 12. pp. 2796-808.

Bibtex

@article{90347d60eb0011deba73000ea68e967b,
title = "Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics",
abstract = "Aberrant signaling causes many diseases, and manipulating signaling pathways with kinase inhibitors has emerged as a promising area of drug research. Most kinase inhibitors target the conserved ATP-binding pocket; therefore specificity is a major concern. Proteomics has previously been used to identify the direct targets of kinase inhibitors upon affinity purification from cellular extracts. Here we introduce a complementary approach to evaluate the effects of kinase inhibitors on the entire cell signaling network. We used triple labeling SILAC (stable isotope labeling by amino acids in cell culture) to compare cellular phosphorylation levels for control, epidermal growth factor stimulus, and growth factor combined with kinase inhibitors. Of thousands of phosphopeptides, less than 10% had a response pattern indicative of targets of U0126 and SB202190, two widely used MAPK inhibitors. Interestingly, 83% of the growth factor-induced phosphorylation events were affected by either or both inhibitors, showing quantitatively that early signaling processes are predominantly transmitted through the MAPK cascades. In contrast to MAPK inhibitors, dasatinib, a clinical drug directed against BCR-ABL, which is the cause of chronic myelogenous leukemia, affected nearly 1,000 phosphopeptides. In addition to the proximal effects on ABL and its immediate targets, dasatinib broadly affected the downstream MAPK pathways. Pathway mapping of regulated sites implicated a variety of cellular functions, such as chromosome remodeling, RNA splicing, and cytoskeletal organization, some of which have been described in the literature before. Our assay is streamlined and generic and could become a useful tool in kinase drug development.",
author = "Cuiping Pan and Olsen, {Jesper V} and Henrik Daub and Matthias Mann",
year = "2009",
doi = "10.1074/mcp.M900285-MCP200",
language = "English",
volume = "8",
pages = "2796--808",
journal = "Molecular and Cellular Proteomics",
issn = "1535-9476",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "12",

}

RIS

TY - JOUR

T1 - Global effects of kinase inhibitors on signaling networks revealed by quantitative phosphoproteomics

AU - Pan, Cuiping

AU - Olsen, Jesper V

AU - Daub, Henrik

AU - Mann, Matthias

PY - 2009

Y1 - 2009

N2 - Aberrant signaling causes many diseases, and manipulating signaling pathways with kinase inhibitors has emerged as a promising area of drug research. Most kinase inhibitors target the conserved ATP-binding pocket; therefore specificity is a major concern. Proteomics has previously been used to identify the direct targets of kinase inhibitors upon affinity purification from cellular extracts. Here we introduce a complementary approach to evaluate the effects of kinase inhibitors on the entire cell signaling network. We used triple labeling SILAC (stable isotope labeling by amino acids in cell culture) to compare cellular phosphorylation levels for control, epidermal growth factor stimulus, and growth factor combined with kinase inhibitors. Of thousands of phosphopeptides, less than 10% had a response pattern indicative of targets of U0126 and SB202190, two widely used MAPK inhibitors. Interestingly, 83% of the growth factor-induced phosphorylation events were affected by either or both inhibitors, showing quantitatively that early signaling processes are predominantly transmitted through the MAPK cascades. In contrast to MAPK inhibitors, dasatinib, a clinical drug directed against BCR-ABL, which is the cause of chronic myelogenous leukemia, affected nearly 1,000 phosphopeptides. In addition to the proximal effects on ABL and its immediate targets, dasatinib broadly affected the downstream MAPK pathways. Pathway mapping of regulated sites implicated a variety of cellular functions, such as chromosome remodeling, RNA splicing, and cytoskeletal organization, some of which have been described in the literature before. Our assay is streamlined and generic and could become a useful tool in kinase drug development.

AB - Aberrant signaling causes many diseases, and manipulating signaling pathways with kinase inhibitors has emerged as a promising area of drug research. Most kinase inhibitors target the conserved ATP-binding pocket; therefore specificity is a major concern. Proteomics has previously been used to identify the direct targets of kinase inhibitors upon affinity purification from cellular extracts. Here we introduce a complementary approach to evaluate the effects of kinase inhibitors on the entire cell signaling network. We used triple labeling SILAC (stable isotope labeling by amino acids in cell culture) to compare cellular phosphorylation levels for control, epidermal growth factor stimulus, and growth factor combined with kinase inhibitors. Of thousands of phosphopeptides, less than 10% had a response pattern indicative of targets of U0126 and SB202190, two widely used MAPK inhibitors. Interestingly, 83% of the growth factor-induced phosphorylation events were affected by either or both inhibitors, showing quantitatively that early signaling processes are predominantly transmitted through the MAPK cascades. In contrast to MAPK inhibitors, dasatinib, a clinical drug directed against BCR-ABL, which is the cause of chronic myelogenous leukemia, affected nearly 1,000 phosphopeptides. In addition to the proximal effects on ABL and its immediate targets, dasatinib broadly affected the downstream MAPK pathways. Pathway mapping of regulated sites implicated a variety of cellular functions, such as chromosome remodeling, RNA splicing, and cytoskeletal organization, some of which have been described in the literature before. Our assay is streamlined and generic and could become a useful tool in kinase drug development.

U2 - 10.1074/mcp.M900285-MCP200

DO - 10.1074/mcp.M900285-MCP200

M3 - Journal article

C2 - 19651622

VL - 8

SP - 2796

EP - 2808

JO - Molecular and Cellular Proteomics

JF - Molecular and Cellular Proteomics

SN - 1535-9476

IS - 12

ER -

ID: 16330608