Warfarin-resistant mutations weaken the binding of human vitamin K epoxide reductase to warfarin
- VernacularTitle:华法林耐药突变体减弱人维生素K环氧化物还原酶与华法林的结合能力
- Author:
Wenqi WANG
1
;
Yan SHEN
;
Xiaoming LI
;
Guomin SHEN
Author Information
- Keywords: blood coagulation; drug resistance; electrophoretic mobility assay; vitamin K; vitamin K epoxide re-ductase; warfarin-resistant mutants; warfarin; anticoagulant
- From: Acta Universitatis Medicinalis Anhui 2024;59(9):1629-1635
- CountryChina
- Language:Chinese
- Abstract: Objective To understand how warfarin inhibition is hindered by the vitamin K epoxide reductase(VKORC1)resistant mutations.Methods An electrophoretic mobility assay was conducted based on the principle that binding with warfarin resulted in changes in VKORC1 electrophoretic mobility.The activity of VKORC1 and the half maximal inhibitory concentration(ICs0)of warfarin inhibiting VKORC1 were detected by ELISA.These experiments evaluated the binding ability of VKORC1 resistant mutants to warfarin.Results With warfarin bound,VKORC1 was protected from N-ethylmaleimide(NEM)modification and showed increased electrophoretic mobility,which was dependent on an unmodified cysteine on the flexible transmembrane helix 1(TM1)of VKORC1.In-creasing the warfarin concentration could shift more VKORC1 towards the species with the fast mobility.Compared to the wildtype VKORC1,the fast mobility fraction became less or disappeared in warfarin-resistant mutants,indi-cating weakened binding of warfarin.In addition,VKORC1 mutants with the weaker electrophoretic mobility shift indicated the stronger warfarin resistant.Conclusion Weakened warfarin binding is the primary cause of warfarin resistance.
- Full text:202412242114169343华法林耐药突变体减弱人维生...物还原酶与华法林的结合能力_王雯琪.pdf
