Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

Leuk Res. 2006 Sep;30(9):1097-104. doi: 10.1016/j.leukres.2006.01.001. Epub 2006 Feb 7.


STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinogens* / metabolism
  • Cell Line, Tumor
  • Chromatography, Liquid
  • Humans
  • Leukemia, Myeloid, Acute / genetics*
  • Leukemia, Myeloid, Acute / metabolism
  • Mass Spectrometry
  • Mutation
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / metabolism
  • Phosphorylation
  • Protein Processing, Post-Translational / genetics
  • Proteomics


  • Carcinogens
  • Neoplasm Proteins