{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T03:48:04Z","timestamp":1790826484210,"version":"4.1.0"},"reference-count":43,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2020,5,29]],"date-time":"2020-05-29T00:00:00Z","timestamp":1590710400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004329","name":"Javna Agencija za Raziskovalno Dejavnost RS","doi-asserted-by":"publisher","award":["P1-0207"],"award-info":[{"award-number":["P1-0207"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>A procoagulant snake venom serine protease was isolated from the venom of the nose-horned viper (Vipera ammodytes ammodytes). This 34 kDa glycoprotein, termed VaaSP-VX, possesses five kDa N-linked carbohydrates. Amino acid sequencing showed VaaSP-VX to be a chymotrypsin-like serine protease. Structurally, it is highly homologous to VaaSP-6 from the same venom and to nikobin from the venom of Vipera nikolskii, neither of which have known functions. VaaSP-VX does not affect platelets. The specific proteolysis of blood coagulation factors X and V by VaaSP-VX suggests that its blood-coagulation-inducing effect is due to its ability to activate these two blood coagulation factors, which following activation, combine to form the prothrombinase complex. VaaSP-VX may thus represent the first example of a serine protease with such a dual activity, which makes it a highly suitable candidate to replace diluted Russell\u2019s viper venom in lupus anticoagulant testing, thus achieving greater reliability of the analysis. As a blood-coagulation-promoting substance that is resistant to serpin inhibition, VaaSP-VX is also interesting from the therapeutic point of view for treating patients suffering from hemophilia.<\/jats:p>","DOI":"10.3390\/toxins12060358","type":"journal-article","created":{"date-parts":[[2020,6,3]],"date-time":"2020-06-03T04:12:09Z","timestamp":1591157529000},"page":"358","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":51,"title":["The Procoagulant Snake Venom Serine Protease Potentially Having a Dual, Blood Coagulation Factor V and X-Activating Activity"],"prefix":"10.3390","volume":"12","author":[{"given":"Zorica","family":"Latinovi\u0107","sequence":"first","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Jo\u017eef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia"},{"name":"Jo\u017eef Stefan International Postgraduate School, Jamova cesta 39, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9854-9955","authenticated-orcid":false,"given":"Adrijana","family":"Leonardi","sequence":"additional","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Jo\u017eef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0002-4603","authenticated-orcid":false,"given":"Cho Yeow","family":"Koh","sequence":"additional","affiliation":[{"name":"Protein Science Laboratory, Department of Biological Sciences, Faculty of Science, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore"},{"name":"Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, 1E Kent Ridge Road, Singapore 119228, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6100-3251","authenticated-orcid":false,"given":"R. Manjunatha","family":"Kini","sequence":"additional","affiliation":[{"name":"Protein Science Laboratory, Department of Biological Sciences, Faculty of Science, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alenka","family":"Trampu\u0161 Bakija","sequence":"additional","affiliation":[{"name":"Division of Pediatrics, University Medical Center, Bohori\u010deva 20, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u017ee","family":"Punger\u010dar","sequence":"additional","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Jo\u017eef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0203-0708","authenticated-orcid":false,"given":"Igor","family":"Kri\u017eaj","sequence":"additional","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Jo\u017eef Stefan Institute, Jamova cesta 39, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4544","DOI":"10.1111\/j.1742-4658.2011.08115.x","article-title":"Enzymatic toxins from snake venom: Structural characterization and mechanism of catalysis","volume":"278","author":"Kang","year":"2011","journal-title":"FEBS J."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1016\/j.toxicon.2011.01.006","article-title":"Haemostatically active proteins in snake venoms","volume":"57","author":"Sajevic","year":"2011","journal-title":"Toxicon"},{"key":"ref_3","unstructured":"Gopalakrishnakone, P., Inagaki, H., Vogel, C.W., Mukherjee, A.K., and Rahmy, T.R. (2017). Snake venom components affecting the coagulation system. Snake Venoms, Springer."},{"key":"ref_4","first-page":"225","article-title":"Snake venom activators of factor X: An overview","volume":"31","author":"Tans","year":"2001","journal-title":"Haemostasis"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1093\/nar\/gkr987","article-title":"Merops: The database of proteolytic enzymes, their substrates and inhibitors","volume":"40","author":"Rawlings","year":"2012","journal-title":"Nucleic Acids Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1016\/S0969-2126(98)00119-1","article-title":"The crystal structure of the novel snake venom plasminogen activator TSV-PA: A prototype structure for snake venom serine proteinases","volume":"6","author":"Parry","year":"1998","journal-title":"Structure"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.biochi.2010.10.004","article-title":"A new tyrosine-specific chymotrypsin-like and angiotensin-degrading serine proteinase from Vipera lebetina snake venom","volume":"93","author":"Siigur","year":"2011","journal-title":"Biochimie"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1016\/j.toxicon.2005.02.020","article-title":"Snake venom serine proteinases: Sequence homology vs. substrate specificity, a paradox to be solved","volume":"45","author":"Serrano","year":"2005","journal-title":"Toxicon"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.biocel.2018.09.011","article-title":"Snake venom components in medicine: From the symbolic rod of Asclepius to tangible medical research and application","volume":"104","author":"Johanningmeier","year":"2018","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"714218","DOI":"10.1155\/2014\/714218","article-title":"Advances in medical revascularisation treatments in acute ischemic stroke","volume":"2014","author":"Asadi","year":"2014","journal-title":"Thrombosis"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1874","DOI":"10.2174\/0929867324666170605091546","article-title":"Snake Venom: From Deadly Toxins to Life-saving Therapeutics","volume":"24","author":"Waheed","year":"2017","journal-title":"Curr. Med. Chem."},{"key":"ref_12","first-page":"191","article-title":"Venomous snakebites in southern Croatia","volume":"30","author":"Prgomet","year":"2006","journal-title":"Coll. Antropol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.toxicon.2016.09.001","article-title":"Cardiotoxic effects of the Vipera ammodytes ammodytes venom fractions in the isolated perfused rat heart","volume":"121","author":"Karabuva","year":"2016","journal-title":"Toxicon"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"33","DOI":"10.3109\/15569543.2013.835827","article-title":"An overview of hemostatically active components of Vipera ammodytes ammodytes venom","volume":"33","author":"Sajevic","year":"2014","journal-title":"Toxin Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1042\/bj1530409","article-title":"Purification and properties of a kininogenin from the venom of Vipera ammodytes ammodytes","volume":"153","author":"Bailey","year":"1976","journal-title":"Biochem. J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.toxicon.2013.11.007","article-title":"VaSP1, catalytically active serine proteinase from Vipera ammodytes ammodytes venom with unconventional active site triad","volume":"77","author":"Brgles","year":"2014","journal-title":"Toxicon"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2287","DOI":"10.1021\/acs.jproteome.9b00120","article-title":"Comprehensive study of the proteome and transcriptome of the venom of the most venomous european viper: Discovery of a new subclass of ancestral snake venom metalloproteinase precursor-derived proteins","volume":"18","author":"Leonardi","year":"2019","journal-title":"J. Proteome Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11233","DOI":"10.1016\/S0021-9258(18)60949-4","article-title":"Proteolysis of factor Va by factor Xa and activated protein C","volume":"262","author":"Odegaard","year":"1987","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2971","DOI":"10.1016\/S0021-9258(19)50682-2","article-title":"The complete cDNA sequence of bovine coagulation factor V","volume":"267","author":"Guinto","year":"1992","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"18614","DOI":"10.1074\/jbc.M007134200","article-title":"The role of the membrane in the inactivation of factor Va by plasmin. Amino acid region 307-348 of factor V plays a critical role in factor Va cofactor function","volume":"276","author":"Kalafatis","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_21","first-page":"374","article-title":"Molecular cloning and analysis of cDNA sequences encoding serine proteinase and Kunitz type inhibitor in venom gland of Vipera nikolskii viper","volume":"37","author":"Ramazanova","year":"2011","journal-title":"Bioorg. Khim."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.toxicon.2007.02.018","article-title":"BJ-48, a novel thrombin-like enzyme from the Bothrops jararacussu venom with high selectivity for Arg over Lys in P1: Role of N-glycosylation in thermostability and active site accessibility","volume":"50","author":"Guedes","year":"2007","journal-title":"Toxicon"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1476","DOI":"10.1016\/j.biochi.2013.03.015","article-title":"Coagulant thrombin-like enzyme (barnettobin) from Bothrops barnetti venom: Molecular sequence analysis of its cDNA and biochemical properties","volume":"95","author":"Sandoval","year":"2013","journal-title":"Biochimie"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1007\/s00204-012-0957-5","article-title":"Crystal structure and activating effect on RyRs of AhV-TL-I, a glycosylated thrombin-like enzyme from Agkistrodon halys snake venom","volume":"87","author":"Zeng","year":"2013","journal-title":"Arch. Toxicol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.ijbiomac.2014.02.058","article-title":"Characterization of a Kunitz-type protease inhibitor peptide (Rusvikunin) purified from Daboia russelii russelii venom","volume":"67","author":"Mukherjee","year":"2014","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"515","DOI":"10.4103\/0019-5049.144643","article-title":"Overview of the coagulation system","volume":"58","author":"Palta","year":"2014","journal-title":"Indian J. Anaesth."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1951","DOI":"10.1111\/j.1538-7836.2009.03622.x","article-title":"The molecular basis of factor V and VIII procofactor activation","volume":"7","author":"Camire","year":"2009","journal-title":"J. Thromb. Haemost."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1111\/j.1432-1033.1997.00012.x","article-title":"Cleavage requirements for activation of factor V by factor Xa","volume":"247","author":"Thorelli","year":"1997","journal-title":"Eur. J. Biochem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"17471","DOI":"10.1016\/S0021-9258(19)77860-0","article-title":"The factor V-activating enzyme (RVV-V) from Russell\u2019s viper venom. Identification of isoproteins RVV-V alpha, -V beta, and -V gamma and their complete amino acid sequences","volume":"263","author":"Tokunaga","year":"1988","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/S0167-4838(98)00232-5","article-title":"Isolation, properties and N-terminal amino acid sequence of a factor V activator from Vipera lebetina (Levantine viper) snake venom","volume":"1429","author":"Siigur","year":"1998","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1002\/prot.21051","article-title":"Structural models of the snake venom factor V activators from Daboia russelli and Daboia lebetina","volume":"64","author":"Segers","year":"2006","journal-title":"Proteins"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1190","DOI":"10.1016\/S0006-3495(02)75476-3","article-title":"Structure and dynamics of zymogen human blood coagulation factor X","volume":"82","author":"Venkateswarlu","year":"2002","journal-title":"Biophys. J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1016\/0041-0101(95)00077-Y","article-title":"Isolation and properties of a blood coagulation factor X activator from the venom of king cobra (Ophiophagus hanah)","volume":"33","author":"Lee","year":"1995","journal-title":"Toxicon"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1016\/0041-0101(95)00070-3","article-title":"An activator of blood coagulation factor X from the venom of Bungarus fasciatus","volume":"33","author":"Zhang","year":"1995","journal-title":"Toxicon"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmed.2019.00025","article-title":"SERPINs-From Trap to Treatment","volume":"6","author":"Sanrattana","year":"2019","journal-title":"Front. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.toxicon.2011.05.007","article-title":"Effect of purified Russell\u2019s viper venom-factor X activator (RVV-X) on renal hemodynamics, renal functions, and coagulopathy in rats","volume":"58","author":"Suntravat","year":"2011","journal-title":"Toxicon"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1007\/978-1-4939-7196-1_14","article-title":"Lupus anticoagulant testing: Diluted Russell Viper Venom Time (dRVVT)","volume":"1646","author":"Pengo","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1006\/jaut.2000.0414","article-title":"Use of the dilute Russell viper venom time (dRVVT): Its importance and pitfalls","volume":"15","author":"Triplett","year":"2000","journal-title":"J. Autoimmun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1055\/s-0038-1669785","article-title":"The first intrinsic tenase complex ihibitor with serine protease structure offers a new perspective in anticoagulant therapy","volume":"118","author":"Leonardi","year":"2018","journal-title":"Thromb. Haemost."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5046","DOI":"10.1021\/pr3006155","article-title":"Conus consors snail venom proteomics proposes functions, pathways, and novel families involved in its venomic system","volume":"11","author":"Leonardi","year":"2012","journal-title":"J. Proteome Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.toxicon.2013.11.009","article-title":"Hemorrhagin VaH4, a covalent heterodimeric P-III metalloproteinase from Vipera ammodytes ammodytes with a potential antitumour activity","volume":"77","author":"Leonardi","year":"2014","journal-title":"Toxicon"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.toxicon.2008.07.015","article-title":"Two coagulation factor X activators from Vipera a. ammodytes venom with potential to treat patients with dysfunctional factors IXa or VIIa","volume":"52","author":"Leonardi","year":"2008","journal-title":"Toxicon"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1158","DOI":"10.1016\/j.biochi.2013.01.003","article-title":"VaH3, one of the principal hemorrhagins in Vipera ammodytes ammodytes venom, is a homodimeric P-IIIc metalloproteinase","volume":"95","author":"Sajevic","year":"2013","journal-title":"Biochimie"}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/6\/358\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:33:49Z","timestamp":1760175229000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/12\/6\/358"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5,29]]},"references-count":43,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2020,6]]}},"alternative-id":["toxins12060358"],"URL":"https:\/\/doi.org\/10.3390\/toxins12060358","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5,29]]}}}