Essential Role of Enzymatic Activity in the Leishmanicidal Mechanism of the Eosinophil Cationic Protein (RNase 3)
Abengozar, María Ángeles (Centro de Investigaciones Biológicas (Madrid))
Fernández-Reyes, María (Centro de Investigaciones Biológicas (Madrid))
Salazar, V. A (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Torrent Burgas, Marc (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
de la Torre, Beatriz G. (Universitat Pompeu Fabra)
Andreu Martínez, David (Universitat Pompeu Fabra)
Boix i Borràs, Esther (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Rivas, Luis (Centro de Investigaciones Biológicas (Madrid))

Data:	2022
Resum:	The recruitment of eosinophils into Leishmania lesions is frequently associated with a favorable evolution. A feasible effector for this process is eosinophil cationic protein (ECP, RNase 3), one of the main human eosinophil granule proteins, endowed with a broad spectrum of antimicrobial activity, including parasites. ECP was active on Leishmania promastigotes and axenic amastigotes (LC's = 3 and 16 μM, respectively) but, in contrast to the irreversible membrane damage caused on bacteria and reproduced by its N -terminal peptides, it only induced a mild and transient plasma membrane destabilization on Leishmania donovani promastigotes. To assess the contribution of RNase activity to the overall leishmanicidal activity of ECP, parasites were challenged in parallel with a single-mutant version, ECP-H15A, devoid of RNase activity, that fully preserves the conformation and liposome permeabilization ability. ECP-H15A showed a similar uptake to ECP on promastigotes, but with higher LC's (>25 μM) for both parasite stages. ECP-treated promastigotes showed a degraded RNA pattern, absent in ECP-H15A-treated samples. Moreover ECP, but not ECP-H15A, reduced more than 2-fold the parasite burden of infected macrophages. Altogether, our results suggest that ECP enters the Leishmania cytoplasm by an endocytic pathway, ultimately leading to RNA degradation as a key contribution to the leishmanicidal mechanism. Thus, ECP combines both membrane destabilization and enzymatic activities to effect parasite killing. Taken together, our data highlight the microbicidal versatility of ECP as an innate immunity component and support the development of cell-penetrating RNases as putative leishmanicidal agents.
Ajuts:	Agencia Estatal de Investigación PID2019-108166GB-I00 Ministerio de Ciencia e Innovación SAF2011-24899 Ministerio de Economía y Competitividad RD16/0027/0010 Agència de Gestió d'Ajuts Universitaris i de Recerca 2009/SGR-492
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:	Membrane disruption ; Cell-penetrating enzyme ; Antimicrobial peptide ; RNase ; Protozoa
Publicat a:	ACS infectious diseases, Vol. 8, Issue 7 (July 2022) , p. 1207-1217, ISSN 2373-8227

DOI: 10.1021/acsinfecdis.1c00537
PMID: 35731709

11 p, 5.2 MB

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