Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry

Hui Min Zhang, Xiu Yu, Michael J. Greig, Ketan S. Gajiwala, Joe C. Wu, Wade Diehl, Elizabeth A. Lunney, Mark Emmett, Alan G. Marshall

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Mutations of the receptor tyrosine kinase KIT are linked to certain cancers such as gastrointestinal stromal tumors (GISTs). Biophysical, biochemical, and structural studies have provided insight into the molecular basis of resistance to the KIT inhibitors, imatinib and sunitinib. Here, solution-phase hydrogen/deuterium exchange (HDX) and direct binding mass spectrometry experiments provide a link between static structure models and the dynamic equilibrium of the multiple states of KIT, supporting that sunitinib targets the autoinhibited conformation of WT-KIT. The D816H mutation shifts the KIT conformational equilibrium toward the activated state. The V560D mutant exhibits two low energy conformations: one is more flexible and resembles the D816H mutant shifted toward the activated conformation, and the other is less flexible and resembles the wild-type KIT in the autoinhibited conformation. This result correlates with the V560D mutant exhibiting a sensitivity to sunitinib that is less than for WT KIT but greater than for KIT D816H. These findings support the elucidation of the resistance mechanism for the KIT mutants. Published by Wiley-Blackwell.

Original languageEnglish (US)
Pages (from-to)703-715
Number of pages13
JournalProtein Science
Volume19
Issue number4
DOIs
StatePublished - Apr 2010
Externally publishedYes

Fingerprint

Deuterium
Drug Resistance
Protein-Tyrosine Kinases
Mass spectrometry
Conformations
Hydrogen
Mass Spectrometry
Pharmaceutical Preparations
Mutation
Gastrointestinal Stromal Tumors
Receptor Protein-Tyrosine Kinases
Model structures
Tumors
Ion exchange
sunitinib
Neoplasms
Experiments

Keywords

  • Conformation
  • Drug resistance
  • Hydrogen/deuterium exchange
  • Mass spectrometry
  • Tyrosine kinase

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Zhang, H. M., Yu, X., Greig, M. J., Gajiwala, K. S., Wu, J. C., Diehl, W., ... Marshall, A. G. (2010). Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry. Protein Science, 19(4), 703-715. https://doi.org/10.1002/pro.347

Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry. / Zhang, Hui Min; Yu, Xiu; Greig, Michael J.; Gajiwala, Ketan S.; Wu, Joe C.; Diehl, Wade; Lunney, Elizabeth A.; Emmett, Mark; Marshall, Alan G.

In: Protein Science, Vol. 19, No. 4, 04.2010, p. 703-715.

Research output: Contribution to journalArticle

Zhang, HM, Yu, X, Greig, MJ, Gajiwala, KS, Wu, JC, Diehl, W, Lunney, EA, Emmett, M & Marshall, AG 2010, 'Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry', Protein Science, vol. 19, no. 4, pp. 703-715. https://doi.org/10.1002/pro.347
Zhang, Hui Min ; Yu, Xiu ; Greig, Michael J. ; Gajiwala, Ketan S. ; Wu, Joe C. ; Diehl, Wade ; Lunney, Elizabeth A. ; Emmett, Mark ; Marshall, Alan G. / Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry. In: Protein Science. 2010 ; Vol. 19, No. 4. pp. 703-715.
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