Isolation and Characterization of NpCI, a New Metallocarboxypeptidase Inhibitor from the Marine Snail Nerita peloronta with Anti- Plasmodium falciparum Activity
Cabrera-Muñoz, Aymara (Universidad de la Habana. Centro de Estudio de Proteínas)
Sierra-Gómez, Yusvel (Universidad de la Habana. Centro de Estudio de Proteínas)
Covaleda Cortés, Giovanni (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Reytor, Mey L. (Universidad de la Habana. Centro de Estudio de Proteínas)
González-González, Yamile (Universidad de la Habana. Centro de Estudio de Proteínas)
Bautista, José M. (Universidad Complutense de Madrid. Departamento de Bioquímica y Biología Molecular)
Avilés, Francesc Xavier (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Alonso del Rivero, Maday (Universidad de la Habana. Centro de Estudio de Proteínas)

Data:	2023
Resum:	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·× 10 −8 mol/L) and porcine CPB (Ki = 8. 15·× 10 −8 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 (IC = 5. 5 μmol/L), reducing parasitaemia mainly by inhibiting the later stages of the parasite's 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's 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.
Ajuts:	Ministerio de Ciencia e Innovación BIO2016-78057-R
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Metallocarboxipeptidases ; Carboxypeptidase inhibitor ; Slow- and tight-binding inhibitor ; Marine invertebrate ; Plasmodium falciparum ; Antimalarial
Publicat a:	Marine drugs, Vol. 21 (january 2023) , ISSN 1660-3397

DOI: 10.3390/md21020094
PMID: 36827135

17 p, 3.1 MB

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