Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target
SUMMARY The proton-translocating inorganic pyrophosphatases (H + -PPases) are primary electrogenic H + pumps that derive energy from the hydrolysis of inorganic pyrophosphate (PPi). They are widely distributed among most land plants and have also been found in several species of protozoan parasites....
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crcambridgeupr:10.1017/s0031182014001267 2024-09-15T18:40:01+00:00 Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target MALLO, NATALIA LAMAS, JESÚS PIAZZON, CARLA LEIRO, JOSÉ M. 2014 http://dx.doi.org/10.1017/s0031182014001267 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182014001267 en eng Cambridge University Press (CUP) https://www.cambridge.org/core/terms Parasitology volume 142, issue 3, page 449-462 ISSN 0031-1820 1469-8161 journal-article 2014 crcambridgeupr https://doi.org/10.1017/s0031182014001267 2024-06-26T04:04:12Z SUMMARY The proton-translocating inorganic pyrophosphatases (H + -PPases) are primary electrogenic H + pumps that derive energy from the hydrolysis of inorganic pyrophosphate (PPi). They are widely distributed among most land plants and have also been found in several species of protozoan parasites. Here we describe, for the first time, the molecular cloning and functional characterization of a gene encoding an H + -pyrophosphatase in the protozoan scuticociliate parasite Philasterides dicentrarchi , which infects turbot. The predicted P. dicentrarchi PPase (PdPPase) consists of 587 amino acids of molecular mass 61·7 kDa and an isoelectric point of 5·0. Several motifs characteristic of plant vacuolar H + -PPases (V–H + -PPases) were also found in the PdPPase, which contains all the sequence motifs of the prototypical type I V–H + -PPase from Arabidopsis thaliana vacuolar pyrophosphatase type I (AVP1) plant. The PdPPase has a characteristic residue that determines strict K + -dependence, but unlike AVP1, PdPPase contains an N-terminal signal peptide (SP) sequence. Antibodies generated by vaccination of mice with a genetic or recombinant protein containing a partial sequence of the PdPPase and a common motif with the polyclonal antibody PAB HK specific to AVP1 recognized a single band of about 62 kDa in western blots. These antibodies specifically stained both vacuole and the alveolar membranes of trophozoites of P. dicentrarchi . H + transport was partially inhibited by the bisphosphonate pamidronate (PAM) and completely inhibited by NaF. The bisphosphonate PAM inhibited both H + -translocation and gene expression. PdPPase and PAM also inhibited in vitro growth of the ciliates. The apparent lack of V–H + -PPases in vertebrates and the parasite sensitivity to PPI analogues may provide a molecular target for developing new drugs to control scuticociliatosis. Article in Journal/Newspaper Turbot Cambridge University Press Parasitology 142 3 449 462 |
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Cambridge University Press |
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English |
description |
SUMMARY The proton-translocating inorganic pyrophosphatases (H + -PPases) are primary electrogenic H + pumps that derive energy from the hydrolysis of inorganic pyrophosphate (PPi). They are widely distributed among most land plants and have also been found in several species of protozoan parasites. Here we describe, for the first time, the molecular cloning and functional characterization of a gene encoding an H + -pyrophosphatase in the protozoan scuticociliate parasite Philasterides dicentrarchi , which infects turbot. The predicted P. dicentrarchi PPase (PdPPase) consists of 587 amino acids of molecular mass 61·7 kDa and an isoelectric point of 5·0. Several motifs characteristic of plant vacuolar H + -PPases (V–H + -PPases) were also found in the PdPPase, which contains all the sequence motifs of the prototypical type I V–H + -PPase from Arabidopsis thaliana vacuolar pyrophosphatase type I (AVP1) plant. The PdPPase has a characteristic residue that determines strict K + -dependence, but unlike AVP1, PdPPase contains an N-terminal signal peptide (SP) sequence. Antibodies generated by vaccination of mice with a genetic or recombinant protein containing a partial sequence of the PdPPase and a common motif with the polyclonal antibody PAB HK specific to AVP1 recognized a single band of about 62 kDa in western blots. These antibodies specifically stained both vacuole and the alveolar membranes of trophozoites of P. dicentrarchi . H + transport was partially inhibited by the bisphosphonate pamidronate (PAM) and completely inhibited by NaF. The bisphosphonate PAM inhibited both H + -translocation and gene expression. PdPPase and PAM also inhibited in vitro growth of the ciliates. The apparent lack of V–H + -PPases in vertebrates and the parasite sensitivity to PPI analogues may provide a molecular target for developing new drugs to control scuticociliatosis. |
format |
Article in Journal/Newspaper |
author |
MALLO, NATALIA LAMAS, JESÚS PIAZZON, CARLA LEIRO, JOSÉ M. |
spellingShingle |
MALLO, NATALIA LAMAS, JESÚS PIAZZON, CARLA LEIRO, JOSÉ M. Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
author_facet |
MALLO, NATALIA LAMAS, JESÚS PIAZZON, CARLA LEIRO, JOSÉ M. |
author_sort |
MALLO, NATALIA |
title |
Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
title_short |
Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
title_full |
Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
title_fullStr |
Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
title_full_unstemmed |
Presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
title_sort |
presence of a plant-like proton-translocating pyrophosphatase in a scuticociliate parasite and its role as a possible drug target |
publisher |
Cambridge University Press (CUP) |
publishDate |
2014 |
url |
http://dx.doi.org/10.1017/s0031182014001267 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182014001267 |
genre |
Turbot |
genre_facet |
Turbot |
op_source |
Parasitology volume 142, issue 3, page 449-462 ISSN 0031-1820 1469-8161 |
op_rights |
https://www.cambridge.org/core/terms |
op_doi |
https://doi.org/10.1017/s0031182014001267 |
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Parasitology |
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142 |
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3 |
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449 |
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462 |
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1810484339677331456 |