Warfarin traps human vitamin K epoxide reductase in an intermediate state during electron transfer

Shen, Guomin; Cui, Weidong; Zhang, Hao; Zhou, Fengbo; Huang, Wei; Liu, Qian; Yang, Yihu; Li, Shuang; Bowman, Gregory R.; Sadler, J. Evan; Gross, Michael L.; Li, Weikai

doi:10.1038/nsmb.3333

Warfarin traps human vitamin K epoxide reductase in an intermediate state during electron transfer

Journal Article · Sun Dec 04 23:00:00 EST 2016 · Nature Structural & Molecular Biology

DOI:https://doi.org/10.1038/nsmb.3333· OSTI ID:1463095

Shen, Guomin ^[1]; Cui, Weidong ^[2]; ^[2]; Zhou, Fengbo ^[3]; Huang, Wei ^[4]; Liu, Qian ^[5]; Yang, Yihu ^[3]; Li, Shuang ^[3]; Bowman, Gregory R. ^[3]; Sadler, J. Evan ^[6]; Gross, Michael L. ^[2]; Li, Weikai ^[3]

Henan Univ. of Science and Technology, Luoyang (China). College of Medicine; Amgen Inc., Cambridge, MA (United States); Washington Univ., St. Louis, MO (United States). School of Medicine and Dept. of Biochemistry and Molecular Biophysics; OSTI
Washington Univ., St. Louis, MO (United States). Dept. of Chemistry
Washington Univ., St. Louis, MO (United States). School of Medicine and Dept. of Biochemistry and Molecular Biophysics
Washington Univ., St. Louis, MO (United States). School of Medicine and Dept. of Biochemistry and Molecular Biophysics; Shanghai Jiao Tong Univ. (China). School of Chemistry and Chemical Engineering
Washington Univ., St. Louis, MO (United States). School of Medicine and Dept. of Biochemistry and Molecular Biophysics; Huazhong Univ. of Science and Technology, Wuhan (China). Tongji Medical College and Dept. of Forensic Medicine
Washington Univ., St. Louis, MO (United States). School of Medicine and Division of Hematology

Although warfarin is the most widely used anticoagulant worldwide, the mechanism by which warfarin inhibits its target, human vitamin K epoxide reductase (hVKOR), remains unclear. We show that warfarin blocks a dynamic electron-transfer process in hVKOR. A major fraction of cellular hVKOR is at an intermediate redox state of this process containing a Cys51-Cys132 disulfide, a characteristic accommodated by a four-transmembrane-helix structure of hVKOR. Warfarin selectively inhibits this major cellular form of hVKOR, whereas disruption of the Cys51-Cys132 disulfide impairs warfarin binding and causes warfarin resistance. Relying on binding interactions identified by cysteine alkylation footprinting and mass spectrometry coupled with mutagenesis analysis, we are able to conduct structure simulations to reveal a closed warfarin-binding pocket stabilized by the Cys51-Cys132 linkage. Understanding the selective warfarin inhibition of a specific redox state of hVKOR should enable the rational design of drugs that exploit the redox chemistry and associated conformational changes in hVKOR.

View Accepted Manuscript (DOE)

Research Organization:: Washington Univ., St. Louis, MO (United States)

Sponsoring Organization:: USDOE; China Scholarship Council (CSC); Burroughs Wellcome Fund (BWF); National Inst. of Health (NIH); American Heart Association (AHA); American Society of Hematology (ASH)

Contributing Organization:: Univ. of North Carolina, Chapel Hill, NC (United States); Umeå Univ. (Sweden)

Grant/Contract Number:: SC0001035

OSTI ID:: 1463095

Journal Information:: Nature Structural & Molecular Biology, Journal Name: Nature Structural & Molecular Biology Journal Issue: 1 Vol. 24; ISSN 1545-9993

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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