{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T15:14:44Z","timestamp":1772118884839,"version":"3.50.1"},"reference-count":25,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2021,11,17]],"date-time":"2021-11-17T00:00:00Z","timestamp":1637107200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100005851","name":"Shahid Beheshti University of Medical Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100005851","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002660","name":"Islamic Azad University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100002660","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteins"],"published-print":{"date-parts":[[2022,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Ecarin is one of the most widely used drug compounds in blood clotting experiments and is used to monitor and treat many diseases such as cancer, liver, lupus, and cardiovascular disease. The metalloproteinase domain is known as the active site of ecarin. In this study, an ecarin metalloproteinase cassette was designed and synthesized in the pUC57 vector. The gene fragment was released and cloned into the pET\u201028a vector and expressed in\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    . The recombinant protein was confirmed by western blotting. Enzyme activity was estimated by a laboratory coagulation test, and prothrombin time and tertiary structure were determined by using the Iterative Threading ASSEmbly Refinement (I\u2010TASSER) server. Data from blood clotting tests for the produced ecarin activity were analyzed using an independent\n                    <jats:italic>t<\/jats:italic>\n                    test. As per I\u2010TASSER server prediction, model 1 with the highest confidence score 0.95, template modeling score (0.84\u2009\u00b1\u20090.08), and root mean square deviation (3.5\u2009\u00b1\u20092.4\u00a0\u00c5) was considered as the best model, and the 2e3xA enzyme was more similar to the target protein. The predictive results helped to better understand the relationship between the structure and function of the ecarin metalloproteinase domain. Also, the production of this active site in the prokaryotic expression system, which is simpler and more cost\u2010effective than the production of the eukaryotic system, showed that this recombinant ecarin could be used as a substitute for the raw snake venom\n                    <jats:italic>of Echis carinatus<\/jats:italic>\n                    because it converts prothrombin into thrombin, and its activity, as estimated using the prothrombin time test, was found to be faster than normal ecarin.\n                  <\/jats:p>","DOI":"10.1002\/prot.26275","type":"journal-article","created":{"date-parts":[[2021,11,5]],"date-time":"2021-11-05T09:29:51Z","timestamp":1636104591000},"page":"802-809","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Prokaryotic expression, evaluation, and prediction of the structure and function of the ecarin metalloproteinase domain"],"prefix":"10.1002","volume":"90","author":[{"given":"Nasrin","family":"Mohammadi","sequence":"first","affiliation":[{"name":"Department of Biology, Science and Research Branch Islamic Azad University  Tehran Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mojgan","family":"Bandehpour","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Biology Research Center Shahid Beheshti University of Medical Sciences  Tehran Iran"},{"name":"Department of Biotechnology, School of Medicine Shahid Beheshti University of Medical Sciences  Tehran Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fattah","family":"Sotoodehnejadnematalahi","sequence":"additional","affiliation":[{"name":"Department of Biology, Science and Research Branch Islamic Azad University  Tehran Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3072-8831","authenticated-orcid":false,"given":"Bahram","family":"Kazemi","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Biology Research Center Shahid Beheshti University of Medical Sciences  Tehran Iran"},{"name":"Department of Biotechnology, School of Medicine Shahid Beheshti University of Medical Sciences  Tehran Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,11,17]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a002982"},{"key":"e_1_2_8_3_1","unstructured":"YonemuraH ImamuraT NakatakeH SoejimaK NozakiC inventors; Chemo Sero Therapeutic Research Institute Kaketsuken assignee.Genetically modified ecarin and process for producing the same. United States patent application US 10\/482 925.2005"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.blre.2019.05.006"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.201000055"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10930\u2010012\u20109409\u20106"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0268-960X(93)90004-N"},{"issue":"22","key":"e_1_2_8_8_1","first-page":"1","article-title":"Current controversies in lupus anticoagulant detection","volume":"5","author":"Moore GW","year":"2016","journal-title":"Antibodies"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0049-3848(76)90026-8"},{"key":"e_1_2_8_10_1","unstructured":"OlsenDR PriorJ SehlLC WallaceDG inventors Cohesion Technologies Inc assignee;Ecarin prothrombin protease and methods. United States patent US 6 413 737.2002."},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.4161\/bioe.22676"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/0049-3848(80)90220-0"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00005a034"},{"key":"e_1_2_8_14_1","unstructured":"https:\/\/www.novoprolabs.com\/tools\/codon-optimization"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1993.1346"},{"key":"e_1_2_8_16_1","unstructured":"https:\/\/zhanglab.ccmb.med.umich.edu\/I-TASSER"},{"key":"e_1_2_8_17_1","doi-asserted-by":"crossref","unstructured":"Igarashi T. Takeda S.2007Crystal structure of Russell's viper venom metalloproteinase.https:\/\/www.rcsb.org\/structure\/2e3x","DOI":"10.2210\/pdb2e3x\/pdb"},{"key":"e_1_2_8_18_1","unstructured":"https:\/\/proteopedia.org\/wiki\/index.php\/2e3x"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0049-3848(75)90150-4"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.25852"},{"key":"e_1_2_8_21_1","unstructured":"McClain S. M.Valasek A.Gruber Coordinated coexpression of thrombin.2016 Google Patents"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.3109\/07388551.2015.1084264"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2009.01.008"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.4161\/bioe.22676"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2007.11.062"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11010\u2010007\u20109603\u20106"}],"container-title":["Proteins: Structure, Function, and Bioinformatics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/prot.26275","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/prot.26275","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/prot.26275","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T06:03:51Z","timestamp":1693202631000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/prot.26275"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,11,17]]},"references-count":25,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2022,3]]}},"alternative-id":["10.1002\/prot.26275"],"URL":"https:\/\/doi.org\/10.1002\/prot.26275","archive":["Portico"],"relation":{"has-review":[{"id-type":"doi","id":"10.1002\/PROT.26275\/v2\/decision1","asserted-by":"object"},{"id-type":"doi","id":"10.1002\/PROT.26275\/v1\/review1","asserted-by":"object"},{"id-type":"doi","id":"10.1002\/PROT.26275\/v1\/decision1","asserted-by":"object"},{"id-type":"doi","id":"10.1002\/PROT.26275\/v3\/decision1","asserted-by":"object"},{"id-type":"doi","id":"10.1002\/PROT.26275\/v2\/review1","asserted-by":"object"}]},"ISSN":["0887-3585","1097-0134"],"issn-type":[{"value":"0887-3585","type":"print"},{"value":"1097-0134","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,11,17]]},"assertion":[{"value":"2021-08-28","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-10-21","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-11-17","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}