{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T10:06:39Z","timestamp":1783850799446,"version":"3.55.0"},"reference-count":50,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2019,3,31]],"date-time":"2019-03-31T00:00:00Z","timestamp":1553990400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["13\/15343-0"],"award-info":[{"award-number":["13\/15343-0"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["15\/15364-3"],"award-info":[{"award-number":["15\/15364-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002322","name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior","doi-asserted-by":"publisher","award":["1736580"],"award-info":[{"award-number":["1736580"]}],"id":[{"id":"10.13039\/501100002322","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Although omics studies have indicated presence of proteases on the Tityus serrulatus venom (TsV), little is known about the function of these molecules. The TsV contains metalloproteases that cleave a series of human neuropeptides, including the dynorphin A (1-13) and the members of neuropeptide Y family. Aiming to isolate the proteases responsible for this activity, the metalloserrulase 3 and 4 (TsMS 3 and TsMS 4) were purified after two chromatographic steps and identified by mass spectrometry analysis. The biochemical parameters (pH, temperature and cation effects) were determined for both proteases, and the catalytic parameters (Km, kcat, cleavage sites) of TsMS 4 over fluorescent substrate were obtained. The metalloserrulases have a high preference for cleaving neuropeptides but presented different primary specificities. For example, the Leu-enkephalin released from dynorphin A (1-13) hydrolysis was exclusively performed by TsMS 3. Neutralization assays using Butantan Institute antivenoms show that both metalloserrulases were well blocked. Although TsMS 3 and TsMS 4 were previously described through cDNA library studies using the venom gland, this is the first time that both these toxins were purified. Thus, this study represents a step further in understanding the mechanism of scorpion venom metalloproteases, which may act as possible neuropeptidases in the envenomation process.<\/jats:p>","DOI":"10.3390\/toxins11040194","type":"journal-article","created":{"date-parts":[[2019,4,2]],"date-time":"2019-04-02T03:21:26Z","timestamp":1554175286000},"page":"194","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Purification and Biochemical Characterization of TsMS 3 and TsMS 4: Neuropeptide-Degrading Metallopeptidases in the Tityus serrulatus Venom"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9234-1758","authenticated-orcid":false,"given":"Daniela","family":"Cajado-Carvalho","sequence":"first","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiane Castilho Fernandes","family":"da Silva","sequence":"additional","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberto Tadashi","family":"Kodama","sequence":"additional","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7296-5475","authenticated-orcid":false,"given":"Douglas Oscar Ceolin","family":"Mariano","sequence":"additional","affiliation":[{"name":"Biochemistry and Biophysics Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2406-0860","authenticated-orcid":false,"given":"Daniel Carvalho","family":"Pimenta","sequence":"additional","affiliation":[{"name":"Biochemistry and Biophysics Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno","family":"Duzzi","sequence":"additional","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre Kazuo","family":"Kuniyoshi","sequence":"additional","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9582-4699","authenticated-orcid":false,"given":"Fernanda Vieira","family":"Portaro","sequence":"additional","affiliation":[{"name":"Immunochemistry Laboratory, Butantan Institute, S\u00e3o Paulo SP 05503-900, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,3,31]]},"reference":[{"key":"ref_1","unstructured":"SINAN (2018, October 10). 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