Imatinib-Induced Changes in Protein Expression and ATP-Binding Affinities of Kinases in Chronic Myelocytic Leukemia Cells.

Abstract

Kinases are crucial components in numerous cell signaling pathways. Aberrant expression and activation of protein kinases are known to be accompanied by many types of cancer, and more than 30 small-molecule kinase inhibitors have been approved by the Food and Drug Administration (FDA) for cancer chemotherapy. Biological and clinical applications of small-molecule kinase inhibitors require comprehensive characterizations about how these inhibitors modulate the protein expression and activities of kinases at the entire proteome scale. In this study, we developed a parallel-reaction monitoring (PRM)-based targeted proteomic method to monitor the alterations in protein expression of kinases in K-562 chronic myelocytic leukemia (CML) cells elicited by treatment with imatinib, an ABL kinase inhibitor approved by the FDA for CML treatment. By employing isotope-coded ATP affinity probes together with liquid chromatography-multiple-reaction monitoring (LC-MRM) analysis, we also examined the modulation of the ATP-binding affinities of kinases induced by imatinib treatment. The results revealed profound increases in protein expression levels of a large number of kinases in K-562 cells upon treatment with imatinib, which is accompanied by substantial decreases in ATP-binding capacities of many kinases. Apart from ABL kinases, we identified a number of other kinases whose ATP-binding affinities are markedly diminished upon imatinib treatment, including CHK1, a checkpoint kinase involved in DNA damage response signaling. Together, our targeted quantitative proteomic methods enabled, for the first time, dual assessments of small-molecule kinase inhibitor-induced changes in protein expression and ATP-binding affinities of kinases in live cells.

• Authors: Weili Miao, Lei Guo, Yinsheng Wang
• Journal: Analytical chemistry (Anal Chem) Vol. 91 Issue 5 Pg. 3209-3214 (03 05 2019) ISSN: 1520-6882 [Electronic] United States
• PMID: 30773012 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Protein Kinase Inhibitors
• Imatinib Mesylate
• Adenosine Triphosphate
• Protein Kinases
• Proto-Oncogene Proteins c-abl

Topics

• Adenosine Triphosphate (chemistry, metabolism)
• DNA Damage (drug effects)
• Humans
• Imatinib Mesylate (pharmacology, therapeutic use)
• K562 Cells
• Leukemia, Myelogenous, Chronic, BCR-ABL Positive (drug therapy, pathology)
• Protein Binding
• Protein Biosynthesis (drug effects)
• Protein Kinase Inhibitors (pharmacology, therapeutic use)
• Protein Kinases (chemistry, metabolism)
• Proteomics (methods)
• Proto-Oncogene Proteins c-abl (chemistry, metabolism)