{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T23:56:30Z","timestamp":1780617390026,"version":"3.54.1"},"reference-count":53,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2022,3,25]],"date-time":"2022-03-25T00:00:00Z","timestamp":1648166400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Bitiscetin-1 (aka bitiscetin) and bitiscetin-2 are C-type lectin-like proteins purified from the venom of Bitis arietans (puff adder). They bind to von Willebrand factor (VWF) and\u2014at least bitiscetin-1\u2014induce platelet agglutination via enhancement of VWF binding to platelet glycoprotein Ib (GPIb). Bitiscetin-1 and -2 bind the VWF A1 and A3 domains, respectively. The A3 domain includes the major site of VWF for binding collagen, explaining why bitiscetin-2 blocks VWF-to-collagen binding. In the present study, sequences for a novel bitiscetin protein\u2014bitiscetin-3\u2014were identified in cDNA constructed from the B. arietans venom gland. The deduced amino acid sequences of bitiscetin-3 subunits \u03b1 and \u03b2 share 79 and 80% identity with those of bitiscetin-1, respectively. Expression vectors for bitiscetin-3\u03b1 and -3\u03b2 were co-transfected to 293T cells, producing the heterodimer protein recombinant bitiscetin-3 (rBit-3). Functionally, purified rBit-3 (1) induced platelet agglutination involving VWF and GPIb, (2) did not compete with bitiscetin-1 for binding to VWF, (3) blocked VWF-to-collagen binding, and (4) lost its platelet agglutination inducing ability in the presence of an anti-VWF monoclonal antibody that blocked VWF-to-collagen binding. These combined results suggest that bitiscetin-3 binds to the A3 domain, as does bitiscetin-2. Except for a small N-terminal fragment of a single subunit\u2014which differs from that of both bitiscetin-3 subunits\u2014the sequences of bitiscetin-2 have never been determined. Therefore, by identifying and analyzing bitiscetin-3, the present study is the first to present the full-length \u03b1- and \u03b2-subunit sequences and recombinant expression of a bitiscetin-family toxin that blocks the binding of VWF to collagen.<\/jats:p>","DOI":"10.3390\/toxins14040236","type":"journal-article","created":{"date-parts":[[2022,3,27]],"date-time":"2022-03-27T21:31:25Z","timestamp":1648416685000},"page":"236","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Bitiscetin-3, a Novel C-Type Lectin-like Protein Cloned from the Venom Gland of the Viper Bitis arietans, Induces Platelet Agglutination and Inhibits Binding of Von Willebrand Factor to Collagen"],"prefix":"10.3390","volume":"14","author":[{"given":"Youhei","family":"Nashimoto","sequence":"first","affiliation":[{"name":"Graduate School of Medical Sciences, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fumio","family":"Matsushita","sequence":"additional","affiliation":[{"name":"Graduate School of Medical Sciences, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes M.","family":"Dijkstra","sequence":"additional","affiliation":[{"name":"Institute for Comprehensive Medical Science, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Nakamura","sequence":"additional","affiliation":[{"name":"Graduate School of Medical Sciences, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidehiko","family":"Akiyama","sequence":"additional","affiliation":[{"name":"Graduate School of Medical Sciences, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiharu","family":"Hamako","sequence":"additional","affiliation":[{"name":"Faculty of Medical Management and Information Science, Fujita Health University School of Health Sciences, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Morita","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Meiji Pharmaceutical University, 2-522-1 Noshio-cho, Kyose 204-8588, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satohiko","family":"Araki","sequence":"additional","affiliation":[{"name":"Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8602, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taei","family":"Matsui","sequence":"additional","affiliation":[{"name":"Graduate School of Medical Sciences, Fujita Health University, 1-98 Kutsukake-cho, Toyoake 470-1192, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1515","DOI":"10.1042\/BCJ20190255","article-title":"Toxins for decoding interface selectivity in nicotinic acetylcholine receptors","volume":"476","author":"Kini","year":"2019","journal-title":"Biochem. 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