{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:49:23Z","timestamp":1760147363340,"version":"build-2065373602"},"reference-count":53,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,28]],"date-time":"2023-01-28T00:00:00Z","timestamp":1674864000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008441","name":"Fundaci\u00f3n Carolina","doi-asserted-by":"publisher","award":["210RT0390","BIO2016-78057-R"],"award-info":[{"award-number":["210RT0390","BIO2016-78057-R"]}],"id":[{"id":"10.13039\/501100008441","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n","award":["210RT0390","BIO2016-78057-R"],"award-info":[{"award-number":["210RT0390","BIO2016-78057-R"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Marine Drugs"],"abstract":"<jats:p>Metallocarboxypeptidases are zinc-dependent peptide-hydrolysing enzymes involved in several important physiological and pathological processes. They have been a target of growing interest in the search for natural or synthetic compound binders with biomedical and drug discovery purposes, i.e., with potential as antimicrobials or antiparasitics. Given that marine resources are an extraordinary source of bioactive molecules, we screened marine invertebrates for new inhibitory compounds with such capabilities. In this work, we report the isolation and molecular and functional characterization of NpCI, a novel strong metallocarboxypeptidase inhibitor from the marine snail Nerita peloronta. NpCI was purified until homogeneity using a combination of affinity chromatography and RP-HPLC. It appeared as a 5921.557 Da protein with 53 residues and six disulphide-linked cysteines, displaying a high sequence similarity with NvCI, a carboxypeptidase inhibitor isolated from Nerita versicolor, a mollusc of the same genus. The purified inhibitor was determined to be a slow- and tight-binding inhibitor of bovine CPA (Ki = 1.1\u00b7\u00d7 10\u22128 mol\/L) and porcine CPB (Ki = 8.15\u00b7\u00d7 10\u22128 mol\/L) and was not able to inhibit proteases from other mechanistic classes. Importantly, this inhibitor showed antiplasmodial activity against Plasmodium falciparum in an in vitro culture (IC50 = 5.5 \u03bcmol\/L), reducing parasitaemia mainly by inhibiting the later stages of the parasite\u2019s intraerythrocytic cycle whilst having no cytotoxic effects on human fibroblasts. Interestingly, initial attempts with other related proteinaceous carboxypeptidase inhibitors also displayed similar antiplasmodial effects. Coincidentally, in recent years, a metallocarboxypeptidase named PfNna1, which is expressed in the schizont phase during the late intraerythrocytic stage of the parasite\u2019s life cycle, has been described. Given that NpCI showed a specific parasiticidal effect on P. falciparum, eliciting pyknotic\/dead parasites, our results suggest that this and related inhibitors could be promising starting agents or lead compounds for antimalarial drug discovery strategies.<\/jats:p>","DOI":"10.3390\/md21020094","type":"journal-article","created":{"date-parts":[[2023,1,30]],"date-time":"2023-01-30T11:19:53Z","timestamp":1675077593000},"page":"94","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Isolation and Characterization of NpCI, a New Metallocarboxypeptidase Inhibitor from the Marine Snail Nerita peloronta with Anti-Plasmodium falciparum Activity"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8208-2446","authenticated-orcid":false,"given":"Aymara","family":"Cabrera-Mu\u00f1oz","sequence":"first","affiliation":[{"name":"Centro de Estudio de Prote\u00ednas, Facultad de Biolog\u00eda, Universidad de la Habana, La Habana 10400, Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yusvel","family":"Sierra-G\u00f3mez","sequence":"additional","affiliation":[{"name":"Centro de Estudio de Prote\u00ednas, Facultad de Biolog\u00eda, Universidad de la Habana, La Habana 10400, Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7621-852X","authenticated-orcid":false,"given":"Giovanni","family":"Covaleda-Cort\u00e9s","sequence":"additional","affiliation":[{"name":"Institut de Biotecnologia i de Biomedicina and Departament de Bioqu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mey L.","family":"Reytor","sequence":"additional","affiliation":[{"name":"Centro de Estudio de Prote\u00ednas, Facultad de Biolog\u00eda, Universidad de la Habana, La Habana 10400, Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yamile","family":"Gonz\u00e1lez-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Centro de Estudio de Prote\u00ednas, Facultad de Biolog\u00eda, Universidad de la Habana, La Habana 10400, Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8926-881X","authenticated-orcid":false,"given":"Jos\u00e9 M.","family":"Bautista","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular, Facultad de Veterinaria, Universidad Complutense de Madrid, Ciudad Universitaria, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1399-6789","authenticated-orcid":false,"given":"Francesc Xavier","family":"Avil\u00e9s","sequence":"additional","affiliation":[{"name":"Institut de Biotecnologia i de Biomedicina and Departament de Bioqu\u00edmica, Universitat Aut\u00f2noma de Barcelona, 08193 Bellaterra, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1988-6852","authenticated-orcid":false,"given":"Maday","family":"Alonso-del-Rivero","sequence":"additional","affiliation":[{"name":"Centro de Estudio de Prote\u00ednas, Facultad de Biolog\u00eda, Universidad de la Habana, La Habana 10400, Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"349","DOI":"10.2174\/138161207780162980","article-title":"Metallocarboxypeptidases: Emerging Drug Targets in Biomedicine","volume":"13","author":"Arolas","year":"2007","journal-title":"Curr. 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