{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:03:12Z","timestamp":1791244992674,"version":"4.1.0"},"reference-count":25,"publisher":"National Academy of Sciences","issue":"51","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2006,12,19]]},"abstract":"<jats:p>More than 21 million prescriptions for warfarin are written yearly in the U.S. Despite its importance, warfarin's target, vitamin K epoxide reductase (VKOR), has resisted purification since its identification in 1972. Here, we report its purification and reconstitution. HPC4, a calcium-specific antibody that recognizes a 12-aa tag, was used to purify and identify VKOR. Partial reconstitution is achieved on the column by washing with 0.4% dioleoylphosphatidylcholine\/0.4% deoxycholate. Activity is completely recovered by dialysis against a buffer containing a reducing agent but lacking dioleoylphosphatidylcholine\/deoxycholate. Removal of detergent from the eluted proteins apparently facilitates liposome formation. Purified recombinant VKOR with tag is \u224821 kDa, as expected; fully active; and &gt;93% pure. The concentration of warfarin for 50% inhibition is the same for purified protein and microsomes. It has been reported that VKOR is a multisubunit enzyme. Our results, however, suggest that a single peptide can accomplish both the conversion of vitamin K epoxide to vitamin K and vitamin K to reduced vitamin K. This purification will allow further characterization of VKOR in relation to other components of the vitamin K cycle and should facilitate its structural determination.<\/jats:p>","DOI":"10.1073\/pnas.0609401103","type":"journal-article","created":{"date-parts":[[2006,12,12]],"date-time":"2006-12-12T22:39:09Z","timestamp":1165963149000},"page":"19308-19313","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":88,"title":["Purified vitamin K epoxide reductase alone is sufficient for conversion of vitamin K epoxide to vitamin K and vitamin K to vitamin KH\n                    <sub>2<\/sub>"],"prefix":"10.1073","volume":"103","author":[{"given":"Pei-Hsuan","family":"Chu","sequence":"first","affiliation":[{"name":"*Department of Biology, University of North Carolina, Chapel Hill, NC 27599-3280;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teng-Yi","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Purdue University, West Lafayette, IN 47907; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason","family":"Williams","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Biology, National Institute on Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, 111 T. 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W.","family":"Stafford","sequence":"additional","affiliation":[{"name":"*Department of Biology, University of North Carolina, Chapel Hill, NC 27599-3280;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2006,12,19]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7569894"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/237032a0"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj1400435"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00601a003"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00346a007"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.46.29068"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.274.34.23963"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature02254"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature02214"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00201a033"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(19)35429-8"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M500765200"},{"key":"e_1_3_4_13_2","first-page":"1","volume":"106","author":"Sun YM","year":"2005","unstructured":"YM Sun, DY Jin, RM Camire, DW Stafford Blood 106, 1\u20135 (2005).","journal-title":"Blood"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2004.04.004"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M413982200"},{"key":"e_1_3_4_16_2","doi-asserted-by":"crossref","first-page":"780","DOI":"10.1160\/TH05-02-0082","volume":"94","author":"Rost S","year":"2005","unstructured":"S Rost, A Fregin, M Hunerberg, CG Bevans, CR Muller, J Oldenburg Thromb Haemost 94, 780\u2013786 (2005).","journal-title":"Thromb Haemost"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2004.05071.x"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M312239200"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M401645200"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-291X(88)81274-9"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M505373200"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi051986y"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.6.2236"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1476-5381.1988.tb11692.x"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1172\/JCI112182"}],"updated-by":[{"DOI":"10.1073\/pnas.1401722111","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2014,2,5]],"date-time":"2014-02-05T00:00:00Z","timestamp":1391558400000}}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.0609401103","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,9]],"date-time":"2023-05-09T19:11:57Z","timestamp":1683659517000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.0609401103"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,12,19]]},"references-count":25,"journal-issue":{"issue":"51","published-print":{"date-parts":[[2006,12,19]]}},"alternative-id":["10.1073\/pnas.0609401103"],"URL":"https:\/\/doi.org\/10.1073\/pnas.0609401103","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,12,19]]},"assertion":[{"value":"2006-08-08","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2006-12-19","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}