Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi
Abstract Proton-translocating inorganic pyrophosphatases (H+-PPases) are an ancient family of membrane bound enzymes that couple pyrophosphate (PPi) hydrolysis to H+ translocation across membranes. In this study, we conducted a molecular characterization of two isoenzymes (PdVP1 and PdVP2) located i...
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ftdoajarticles:oai:doaj.org/article:de441daeb1b04a3b8ac663ec105559f8 2023-05-15T18:41:16+02:00 Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi I. Folgueira J. Lamas R. A. Sueiro J. M. Leiro 2021-04-01T00:00:00Z https://doi.org/10.1038/s41598-021-88102-0 https://doaj.org/article/de441daeb1b04a3b8ac663ec105559f8 EN eng Nature Portfolio https://doi.org/10.1038/s41598-021-88102-0 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-021-88102-0 2045-2322 https://doaj.org/article/de441daeb1b04a3b8ac663ec105559f8 Scientific Reports, Vol 11, Iss 1, Pp 1-15 (2021) Medicine R Science Q article 2021 ftdoajarticles https://doi.org/10.1038/s41598-021-88102-0 2022-12-31T05:19:39Z Abstract Proton-translocating inorganic pyrophosphatases (H+-PPases) are an ancient family of membrane bound enzymes that couple pyrophosphate (PPi) hydrolysis to H+ translocation across membranes. In this study, we conducted a molecular characterization of two isoenzymes (PdVP1 and PdVP2) located in respectively the alveolar sacs and in the membranes of the intracellular vacuoles of a scuticociliate parasite (Philasterides dicentrarchi) of farmed turbot. We analyzed the genetic expression of the isoenzymes after administration of antiparasitic drugs and after infection in the host. PdVP1 and PdVP2 are encoded by two genes of 2485 and 3069 bp, which respectively contain 3 and 11 exons and express proteins of 746 and 810 aa of molecular mass 78.9 and 87.6 kDa. Topological predictions from isoenzyme sequences indicate the formation of thirteen transmembrane regions (TMRs) for PdVP1 and seventeen TMRs for PdVP2. Protein structure modelling indicated that both isoenzymes are homodimeric, with three Mg2+ binding sites and an additional K+ binding site in PdVP2. The levels of identity and similarity between the isoenzyme sequences are respectively 33.5 and 51.2%. The molecular weights of the native proteins are 158 kDa (PdVP1) and 178 kDa (PdVP2). The isoenzyme sequences are derived from paralogous genes that form a monophyletic grouping with other ciliate species. Genetic expression of the isoenzymes is closely related to the acidification of alveolar sacs (PdVP1) and intracellular vacuoles (PdVP2): antiparasitic drugs inhibit transcription, while infection increases transcription of both isoenzymes. The study findings show that P. dicentrarchi possesses two isoenzymes with H+-PPase activity which are located in acidophilic cell compartment membranes and which are activated during infection in the host and are sensitive to antiparasitic drugs. The findings open the way to using molecular modelling to design drugs for the treatment of scuticociliatosis. Article in Journal/Newspaper Turbot Directory of Open Access Journals: DOAJ Articles Scientific Reports 11 1 |
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Medicine R Science Q I. Folgueira J. Lamas R. A. Sueiro J. M. Leiro Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
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Medicine R Science Q |
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Abstract Proton-translocating inorganic pyrophosphatases (H+-PPases) are an ancient family of membrane bound enzymes that couple pyrophosphate (PPi) hydrolysis to H+ translocation across membranes. In this study, we conducted a molecular characterization of two isoenzymes (PdVP1 and PdVP2) located in respectively the alveolar sacs and in the membranes of the intracellular vacuoles of a scuticociliate parasite (Philasterides dicentrarchi) of farmed turbot. We analyzed the genetic expression of the isoenzymes after administration of antiparasitic drugs and after infection in the host. PdVP1 and PdVP2 are encoded by two genes of 2485 and 3069 bp, which respectively contain 3 and 11 exons and express proteins of 746 and 810 aa of molecular mass 78.9 and 87.6 kDa. Topological predictions from isoenzyme sequences indicate the formation of thirteen transmembrane regions (TMRs) for PdVP1 and seventeen TMRs for PdVP2. Protein structure modelling indicated that both isoenzymes are homodimeric, with three Mg2+ binding sites and an additional K+ binding site in PdVP2. The levels of identity and similarity between the isoenzyme sequences are respectively 33.5 and 51.2%. The molecular weights of the native proteins are 158 kDa (PdVP1) and 178 kDa (PdVP2). The isoenzyme sequences are derived from paralogous genes that form a monophyletic grouping with other ciliate species. Genetic expression of the isoenzymes is closely related to the acidification of alveolar sacs (PdVP1) and intracellular vacuoles (PdVP2): antiparasitic drugs inhibit transcription, while infection increases transcription of both isoenzymes. The study findings show that P. dicentrarchi possesses two isoenzymes with H+-PPase activity which are located in acidophilic cell compartment membranes and which are activated during infection in the host and are sensitive to antiparasitic drugs. The findings open the way to using molecular modelling to design drugs for the treatment of scuticociliatosis. |
format |
Article in Journal/Newspaper |
author |
I. Folgueira J. Lamas R. A. Sueiro J. M. Leiro |
author_facet |
I. Folgueira J. Lamas R. A. Sueiro J. M. Leiro |
author_sort |
I. Folgueira |
title |
Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
title_short |
Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
title_full |
Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
title_fullStr |
Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
title_full_unstemmed |
Molecular characterization and transcriptional regulation of two types of H+-pyrophosphatases in the scuticociliate parasite Philasterides dicentrarchi |
title_sort |
molecular characterization and transcriptional regulation of two types of h+-pyrophosphatases in the scuticociliate parasite philasterides dicentrarchi |
publisher |
Nature Portfolio |
publishDate |
2021 |
url |
https://doi.org/10.1038/s41598-021-88102-0 https://doaj.org/article/de441daeb1b04a3b8ac663ec105559f8 |
genre |
Turbot |
genre_facet |
Turbot |
op_source |
Scientific Reports, Vol 11, Iss 1, Pp 1-15 (2021) |
op_relation |
https://doi.org/10.1038/s41598-021-88102-0 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-021-88102-0 2045-2322 https://doaj.org/article/de441daeb1b04a3b8ac663ec105559f8 |
op_doi |
https://doi.org/10.1038/s41598-021-88102-0 |
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Scientific Reports |
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11 |
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1766230760891613184 |