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 language | English (US) |
|---|---|
| Pages (from-to) | 703-715 |
| Number of pages | 13 |
| Journal | Protein Science |
| Volume | 19 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 2010 |
| Externally published | Yes |
Fingerprint
Keywords
- Conformation
- Drug resistance
- Hydrogen/deuterium exchange
- Mass spectrometry
- Tyrosine kinase
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
Cite this
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 journal › Article
}
TY - JOUR
T1 - Drug binding and resistance mechanism of KIT tyrosine kinase revealed by hydrogen/deuterium exchange FTICR mass spectrometry
AU - Zhang, Hui Min
AU - Yu, Xiu
AU - Greig, Michael J.
AU - Gajiwala, Ketan S.
AU - Wu, Joe C.
AU - Diehl, Wade
AU - Lunney, Elizabeth A.
AU - Emmett, Mark
AU - Marshall, Alan G.
PY - 2010/4
Y1 - 2010/4
N2 - 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.
AB - 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.
KW - Conformation
KW - Drug resistance
KW - Hydrogen/deuterium exchange
KW - Mass spectrometry
KW - Tyrosine kinase
UR - http://www.scopus.com/inward/record.url?scp=77950201397&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77950201397&partnerID=8YFLogxK
U2 - 10.1002/pro.347
DO - 10.1002/pro.347
M3 - Article
VL - 19
SP - 703
EP - 715
JO - Protein Science
JF - Protein Science
SN - 0961-8368
IS - 4
ER -