{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T20:25:25Z","timestamp":1780518325805,"version":"3.54.1"},"reference-count":62,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,3,23]],"date-time":"2022-03-23T00:00:00Z","timestamp":1647993600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004329","name":"Slovenian Research Agency","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>Disintegrin-like\/cysteine-rich (DC) proteins have long been regarded just as products of proteolysis of P-III snake venom metalloproteinases (SVMPs). However, here we demonstrate that a DC protein from the venom of Vipera ammodytes (Vaa; nose-horned viper), VaaMPIII-3, is encoded per se by a P-III SVMP-like gene that has a deletion in the region of the catalytic metalloproteinase domain and in part of the non-catalytic disintegrin-like domain. In this way, we justify the proposal of the introduction of a new subclass P-IIIe of SVMP-derived DC proteins. We purified VaaMPIII-3 from the venom of Vaa in a series of chromatographic steps. A covalent chromatography step based on thiol-disulphide exchange revealed that VaaMPIII-3 contains an unpaired Cys residue. This was demonstrated to be Cys6 in about 90% and Cys19 in about 10% of the VaaMPIII-3 molecules. We further constructed a three-dimensional homology model of VaaMPIII-3. From this model, it is evident that both Cys6 and Cys19 can pair with Cys26, which suggests that the intramolecular thiol-disulphide exchange has a regulatory function. VaaMPIII-3 is an acidic 21-kDa monomeric glycoprotein that exists in at least six N-glycoforms, with isoelectric points ranging from pH 4.5 to 5.1. Consistent with the presence of an integrin-binding motif in its sequence, SECD, VaaMPIII-3 inhibited collagen-induced platelet aggregation. It also inhibited ADP- and arachidonic-acid-induced platelet aggregation, but not ristocetin-induced platelet agglutination and the blood coagulation cascade.<\/jats:p>","DOI":"10.3390\/toxins14040232","type":"journal-article","created":{"date-parts":[[2022,3,23]],"date-time":"2022-03-23T22:08:06Z","timestamp":1648073286000},"page":"232","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Genomic Confirmation of the P-IIIe Subclass of Snake Venom Metalloproteinases and Characterisation of Its First Member, a Disintegrin-Like\/Cysteine-Rich Protein"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1233-9864","authenticated-orcid":false,"given":"Kity","family":"Po\u017eek","sequence":"first","affiliation":[{"name":"Department of Molecular and Biomedical Sciences, Jo\u017eef Stefan Institute, 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, 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, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8624-6703","authenticated-orcid":false,"given":"Weiqiao","family":"Rao","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark"},{"name":"Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214128, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zijian","family":"Gao","sequence":"additional","affiliation":[{"name":"Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214128, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Siqi","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Mass Spectrometry, Beijing Genomics Institute-Research, Shenzhen 508083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6918-5574","authenticated-orcid":false,"given":"Andreas Hougaard","family":"Laustsen","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alenka","family":"Trampu\u0161 Bakija","sequence":"additional","affiliation":[{"name":"Division of Pediatrics, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarina","family":"Reber\u0161ek","sequence":"additional","affiliation":[{"name":"Department of Hematology, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3752-2067","authenticated-orcid":false,"given":"Helena","family":"Podgornik","sequence":"additional","affiliation":[{"name":"Department of Hematology, University Medical Centre Ljubljana, SI-1000 Ljubljana, Slovenia"},{"name":"Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 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, SI-1000 Ljubljana, Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,23]]},"reference":[{"key":"ref_1","unstructured":"Mebs, D. 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