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PMID: 20190765 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A chemical and phosphoproteomic characterization of dasatinib action in lung cancer.

Nature chemical biology ·Vol. 6 ·No. 4 ·2010-04-00 ·Pages 291-9

Li J, Rix U, Fang B, Bai Y, Edwards A, Colinge J, Bennett KL, Gao J, Song L, Eschrich S, Superti-Furga G, Koomen J, Haura EB

Abstract

We describe a strategy for comprehending signaling pathways that are active in lung cancer cells and that are targeted by dasatinib using chemical proteomics to identify direct interacting proteins combined with immunoaffinity purification of tyrosine-phosphorylated peptides corresponding to activated tyrosine kinases. We identified nearly 40 different kinase targets of dasatinib. These include SRC-family kinase (SFK) members (LYN, SRC, FYN, LCK and YES), nonreceptor tyrosine kinases (FRK, BRK and ACK) and receptor tyrosine kinases (Ephrin receptors, DDR1 and EGFR). Using quantitative phosphoproteomics, we identified peptides corresponding to autophosphorylation sites of these tyrosine kinases that are inhibited in a concentration-dependent manner by dasatinib. Using drug-resistant gatekeeper mutants, we show that SFKs (particularly SRC and FYN), as well as EGFR, are relevant targets for dasatinib action. The combined mass spectrometry-based approach described here provides a system-level view of dasatinib action in cancer cells and suggests both functional targets and a rationale for combinatorial therapeutic strategies.

MeSH Terms
Apoptosis Cell Line, Tumor Dasatinib Gene Knockdown Techniques Humans Lung Neoplasms/drug therapy,metabolism,pathology Mass Spectrometry Peptides/chemistry,metabolism Phenotype Phosphoproteins/chemistry,metabolism Phosphorylation/drug effects Protein Binding Protein Serine-Threonine Kinases/metabolism Protein-Tyrosine Kinases/antagonists & inhibitors,metabolism Proteomics/methods Pyrimidines/metabolism,pharmacology,therapeutic use Signal Transduction/drug effects Thiazoles/metabolism,pharmacology,therapeutic use
Chemicals
Peptides Phosphoproteins Pyrimidines Thiazoles Protein-Tyrosine Kinases Protein Serine-Threonine Kinases Dasatinib
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Li Jiannong
Department of Thoracic Oncology Program, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA.
Rix Uwe
Fang Bin
Bai Yun
Edwards Arthur
Colinge Jacques
Bennett Keiryn L
Gao Jingchun
Song Lanxi
Eschrich Steven
Superti-Furga Giulio
Koomen John
Haura Eric B
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Article Info
Journal
Nature chemical biology
Abbr.
Nat Chem Biol
ISSN
1552-4469
DOI
10.1038/nchembio.332
Published
2010-04-00
Epub
2010-00-28
Pages
291-9
Language
English
Region
United States
NLM ID
101231976
PMCID
PMC2842457
Subset
IM
Grants
NCI NIH HHS · P50 CA119997 · United States
NCI NIH HHS · R01 CA123174 · United States
NCI NIH HHS · R01 CA123174-02 · United States
Databases
PubChem-Substance
87357382
External Links
PubMed source DOI: 10.1038/nchembio.332 PMC Full Text View journal details
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