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Functional characterization of single-domain cystatin-like cysteine proteinase inhibitors expressed by the trematode Fasciola hepatica

Published online by Cambridge University Press:  12 July 2017

MARTÍN CANCELA ,
ILEANA CORVO ,
EDILEUZA DA SILVA ,
ALINE TEICHMANN ,
LEDA ROCHE ,
ALVARO DÍAZ ,
JOSÉ FRANSISCO TORT ,
HENRIQUE B. FERREIRA  and
ARNALDO ZAHA
MARTÍN CANCELA*
Affiliation:
Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências e Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
ILEANA CORVO
Affiliation:
Departamento de Genética, Facultad de Medicina, Universidad de la República, UDELAR, Montevideo, Uruguay Laboratorio de I+D de Moléculas Bioactivas, CenUR Litoral Norte Sede Paysandú, Universidad de la República, Paysandú, Uruguay
EDILEUZA DA SILVA
Affiliation:
Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências e Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
ALINE TEICHMANN
Affiliation:
Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências e Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
LEDA ROCHE
Affiliation:
Departamento de Genética, Facultad de Medicina, Universidad de la República, UDELAR, Montevideo, Uruguay
ALVARO DÍAZ
Affiliation:
Cátedra de Inmunología, Departamento de Biociencias, Facultad de Química/IQB, Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay
JOSÉ FRANSISCO TORT
Affiliation:
Departamento de Genética, Facultad de Medicina, Universidad de la República, UDELAR, Montevideo, Uruguay
HENRIQUE B. FERREIRA
Affiliation:
Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências e Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
ARNALDO ZAHA
Affiliation:
Departamento de Biologia Molecular e Biotecnologia, Instituto de Biociências e Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
*
*Corresponding author: Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, UFRGS, Caixa Postal 15053, CEP 91501-970, Porto Alegre, RS, Brazil. E-mail: martin@cbiot.ufrgs.br
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Summary

Cystatins are small, phylogenetically conserved proteins that are tight-binding inhibitors of cysteine proteinases. The liver fluke Fasciola hepatica uses a diverse set of cysteine proteinases of the papain superfamily for host invasion, immune evasion and nutrition, but little is known about the regulation of these enzymes. The aim of this work is to characterize the cystatin repertoire of F. hepatica. For this purpose, we first surveyed the available sequence databases, identifying three different F. hepatica single-domain cystatins. In agreement with the in silico predictions, at least three small proteins with cysteine proteinase binding activity were identified. Phylogenetic analyses showed that the three cystatins (named FhStf-1, -2 and -3) are members of the I25A subfamily (stefins). Whereas FhStf-1 grouped with classical stefins, FhStf-2 and 3 fell in a divergent stefin subgroup unusually featuring signal peptides. Recombinant rFhStf-1, -2 and -3 had potent inhibitory activity against F. hepatica cathepsin L cysteine proteinases but differed in their capacity to inhibit mammalian cathepsin B, L and C. FhStf-1 was localized in the F. hepatica reproductive organs (testes and ovary), and at the surface lamella of the adult gut, where it may regulate cysteine proteinases related with reproduction and digestion, respectively. FhStf-1 was also detected among F. hepatica excretion–secretion (E/S) products of adult flukes. This suggests that it is secreted by non-classical secretory pathway and that it may interact with host lysosomal cysteine proteinases.

Keywords

Fasciola hepaticacystatincysteine proteinase regulationparasite invasionparasite–host interaction
Type
Research Article
Information
Parasitology , Volume 144 , Issue 13 , November 2017 , pp. 1695 - 1707
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Copyright
Copyright © Cambridge University Press 2017 

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