@article{876d94e9476e455da8e2160a291f38ef,
title = "Warfarin traps human Vitamin K epoxide reductase in an intermediate state during electron transfer",
abstract = "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. Here we show that warfarin blocks a dynamic electron-transfer process in hVKOR. A major fraction of cellular hVKOR is in an intermediate redox state 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 conducted structure simulations, which revealed 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.",
author = "Guomin Shen and Weidong Cui and Hao Zhang and Fengbo Zhou and Wei Huang and Qian Liu and Yihu Yang and Shuang Li and Bowman, {Gregory R.} and Sadler, {J. Evan} and Gross, {Michael L.} and Weikai Li",
note = "Funding Information: We thank D.W. Stafford, J. Tie, and D. Jin (University of North Carolina) for their gift of double-knockout cell lines used in activity assays and for their generous help in the assay protocol; S. Wanrooij (Ume{\aa} University) for the gift of the T-REx-293 cell line and the protocol for establishing stable cell lines; J. Tie, D.W. Stafford, S. Liu and T. Ellenberger for critical reading of the manuscript. H.Z. is supported by the DOE (DE-SC0001035 to PARC). W.H. is supported by the China Scholarship Council (201206235027). G.R.B. is supported by a BWF Career Award. J.E.S. is supported by the NIH NHLBI (R01 HL130446 and U54 HL112303). M.L.G. is supported by the NIH NIGMS (P41 GM103422), which financed the MS measurements. W.L. is supported by the NHLBI (R01 HL121718), the AHA (grant-in-aid 14GRNT20310017) and an ASH Scholar Award. Publisher Copyright: {\textcopyright} 2017 Nature America, Inc., part of Springer Nature. All rights reserved.",
year = "2017",
month = jan,
day = "5",
doi = "10.1038/nsmb.3333",
language = "English",
volume = "24",
pages = "69--76",
journal = "Nature Structural and Molecular Biology",
issn = "1545-9993",
number = "1",
}