Proteomics analysis of cellular imatinib targets and their candidate downstream effectors

Susanne B. Breitkopf, Felix S. Oppermann, G. Kéri, Markus Grammel, Henrik Daub

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Inhibition of deregulated protein kinases by small molecule drugs has evolved into a major therapeutic strategy for the treatment of human malignancies. Knowledge about direct cellular targets of kinase-selective drugs and the identification of druggable downstream mediators of oncogenic signaling are relevant for both initial therapy selection and the nomination of alternative targets in case molecular resistance emerges. To address these issues, we performed a proof-of-concept proteomics study designed to monitor drug effects on the pharmacologically tractable subproteome isolated by affinity purification with immobilized, nonselective kinase inhibitors. We applied this strategy to chronic myeloid leukemia cells that express the transforming Bcr-Abl fusion kinase. We used SILAC to measure how cellular treatment with the Bcr-Abl inhibitor imatinib affects protein binding to a generic kinase inhibitor resin and further quantified site-specific phosphorylations on resin-retained proteins. Our integrated approach indicated additional imatinib target candidates, such as flavine adenine dinucleotide synthetase, as well as repressed phosphorylation events on downstream effectors not yet implicated in imatinib-regulated signaling. These included activity-regulating phosphorylations on the kinases Btk, Fer, and focal adhesion kinase, which may qualify them as alternative target candidates in Bcr-Abl-driven oncogenesis. Our approach is rather generic and may have various applications in kinase drug discovery.

Original languageEnglish
Pages (from-to)6033-6043
Number of pages11
JournalJournal of Proteome Research
Volume9
Issue number11
DOIs
Publication statusPublished - Nov 5 2010

Fingerprint

Proteomics
Phosphotransferases
Phosphorylation
Resins
Pharmaceutical Preparations
Focal Adhesion Protein-Tyrosine Kinases
Adenine
Myeloid Cells
Drug Discovery
Ligases
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Protein Binding
Protein Kinases
Purification
Imatinib Mesylate
Carcinogenesis
Fusion reactions
Molecules
Therapeutics
Neoplasms

Keywords

  • affinity purification
  • chronic myeloid leukemia
  • imatinib
  • kinase inhibitors
  • phosphoproteomics
  • protein kinases
  • SILAC

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Proteomics analysis of cellular imatinib targets and their candidate downstream effectors. / Breitkopf, Susanne B.; Oppermann, Felix S.; Kéri, G.; Grammel, Markus; Daub, Henrik.

In: Journal of Proteome Research, Vol. 9, No. 11, 05.11.2010, p. 6033-6043.

Research output: Contribution to journalArticle

Breitkopf, Susanne B. ; Oppermann, Felix S. ; Kéri, G. ; Grammel, Markus ; Daub, Henrik. / Proteomics analysis of cellular imatinib targets and their candidate downstream effectors. In: Journal of Proteome Research. 2010 ; Vol. 9, No. 11. pp. 6033-6043.
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