Quantitative Proteomics Analysis of Leukemia Cells

Methods Mol Biol. 2016:1465:139-48. doi: 10.1007/978-1-4939-4011-0_12.

Abstract

Chronic myeloid leukemia (CML) is driven by the oncogenic fusion kinase Bcr-Abl, which organizes its own signaling network with various proteins. These proteins, their interactions, and their role in relevant signaling pathways can be analyzed by quantitative mass spectrometry (MS) approaches in various models systems, e.g., in cell culture models. In this chapter, we describe in detail immunoprecipitations and quantitative proteomics analysis using stable isotope labeling by amino acids in cell culture (SILAC) of components of the Bcr-Abl signaling pathway in the human CML cell line K562.

Keywords: Bcr-Abl; Chronic myeloid leukemia; Gab2; Immunoprecipitation; Mass spectrometry; Posttranslational modification; Protein complex; Protein dynamics; Proteomics; SILAC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Chromatography, Liquid / methods*
  • Humans
  • Immunoprecipitation
  • Isotope Labeling
  • K562 Cells
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / metabolism*
  • Proteomics / methods*
  • Signal Transduction
  • Tandem Mass Spectrometry / methods*