Cargando…
Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development
Infection with Plasmodium falciparum and vivax cause most cases of malaria. Emerging resistance to current antimalarial medications makes new drug development imperative. Ideally a new antimalarial drug should treat both falciparum and vivax malaria. Because malaria parasites are purine auxotrophic,...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4706624/ https://www.ncbi.nlm.nih.gov/pubmed/26862473 http://dx.doi.org/10.1016/j.ijpddr.2015.11.003 |
_version_ | 1782409191891140608 |
---|---|
author | Deniskin, Roman Frame, I.J. Sosa, Yvett Akabas, Myles H. |
author_facet | Deniskin, Roman Frame, I.J. Sosa, Yvett Akabas, Myles H. |
author_sort | Deniskin, Roman |
collection | PubMed |
description | Infection with Plasmodium falciparum and vivax cause most cases of malaria. Emerging resistance to current antimalarial medications makes new drug development imperative. Ideally a new antimalarial drug should treat both falciparum and vivax malaria. Because malaria parasites are purine auxotrophic, they rely on purines imported from the host erythrocyte via Equilibrative Nucleoside Transporters (ENTs). Thus, the purine import transporters represent a potential target for antimalarial drug development. For falciparum parasites the primary purine transporter is the P. falciparum Equilibrative Nucleoside Transporter Type 1 (PfENT1). Recently we identified potent PfENT1 inhibitors with nanomolar IC(50) values using a robust, yeast-based high throughput screening assay. In the current work we characterized the Plasmodium vivax ENT1 (PvENT1) homologue and its sensitivity to the PfENT1 inhibitors. We expressed a yeast codon-optimized PvENT1 gene in Saccharomyces cerevisiae. PvENT1-expressing yeast imported both purines ([(3)H]adenosine) and pyrimidines ([(3)H]uridine), whereas wild type (fui1Δ) yeast did not. Based on radiolabel substrate uptake inhibition experiments, inosine had the lowest IC(50) (3.8 μM), compared to guanosine (14.9 μM) and adenosine (142 μM). For pyrimidines, thymidine had an IC(50) of 183 μM (vs. cytidine and uridine; mM range). IC(50) values were higher for nucleobases compared to the corresponding nucleosides; hypoxanthine had a 25-fold higher IC(50) than inosine. The archetypal human ENT1 inhibitor 4-nitrobenzylthioinosine (NBMPR) had no effect on PvENT1, whereas dipyridamole inhibited PvENT1, albeit with a 40 μM IC(50), a 1000-fold less sensitive than human ENT1 (hENT1). The PfENT1 inhibitors blocked transport activity of PvENT1 and the five known naturally occurring non-synonymous single nucleotide polymorphisms (SNPs) with similar IC(50) values. Thus, the PfENT1 inhibitors also target PvENT1. This implies that development of novel antimalarial drugs that target both falciparum and vivax ENT1 may be feasible. |
format | Online Article Text |
id | pubmed-4706624 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-47066242016-02-09 Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development Deniskin, Roman Frame, I.J. Sosa, Yvett Akabas, Myles H. Int J Parasitol Drugs Drug Resist Article Infection with Plasmodium falciparum and vivax cause most cases of malaria. Emerging resistance to current antimalarial medications makes new drug development imperative. Ideally a new antimalarial drug should treat both falciparum and vivax malaria. Because malaria parasites are purine auxotrophic, they rely on purines imported from the host erythrocyte via Equilibrative Nucleoside Transporters (ENTs). Thus, the purine import transporters represent a potential target for antimalarial drug development. For falciparum parasites the primary purine transporter is the P. falciparum Equilibrative Nucleoside Transporter Type 1 (PfENT1). Recently we identified potent PfENT1 inhibitors with nanomolar IC(50) values using a robust, yeast-based high throughput screening assay. In the current work we characterized the Plasmodium vivax ENT1 (PvENT1) homologue and its sensitivity to the PfENT1 inhibitors. We expressed a yeast codon-optimized PvENT1 gene in Saccharomyces cerevisiae. PvENT1-expressing yeast imported both purines ([(3)H]adenosine) and pyrimidines ([(3)H]uridine), whereas wild type (fui1Δ) yeast did not. Based on radiolabel substrate uptake inhibition experiments, inosine had the lowest IC(50) (3.8 μM), compared to guanosine (14.9 μM) and adenosine (142 μM). For pyrimidines, thymidine had an IC(50) of 183 μM (vs. cytidine and uridine; mM range). IC(50) values were higher for nucleobases compared to the corresponding nucleosides; hypoxanthine had a 25-fold higher IC(50) than inosine. The archetypal human ENT1 inhibitor 4-nitrobenzylthioinosine (NBMPR) had no effect on PvENT1, whereas dipyridamole inhibited PvENT1, albeit with a 40 μM IC(50), a 1000-fold less sensitive than human ENT1 (hENT1). The PfENT1 inhibitors blocked transport activity of PvENT1 and the five known naturally occurring non-synonymous single nucleotide polymorphisms (SNPs) with similar IC(50) values. Thus, the PfENT1 inhibitors also target PvENT1. This implies that development of novel antimalarial drugs that target both falciparum and vivax ENT1 may be feasible. Elsevier 2015-11-28 /pmc/articles/PMC4706624/ /pubmed/26862473 http://dx.doi.org/10.1016/j.ijpddr.2015.11.003 Text en © 2015 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Deniskin, Roman Frame, I.J. Sosa, Yvett Akabas, Myles H. Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title | Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title_full | Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title_fullStr | Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title_full_unstemmed | Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title_short | Targeting the Plasmodium vivax equilibrative nucleoside transporter 1 (PvENT1) for antimalarial drug development |
title_sort | targeting the plasmodium vivax equilibrative nucleoside transporter 1 (pvent1) for antimalarial drug development |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4706624/ https://www.ncbi.nlm.nih.gov/pubmed/26862473 http://dx.doi.org/10.1016/j.ijpddr.2015.11.003 |
work_keys_str_mv | AT deniskinroman targetingtheplasmodiumvivaxequilibrativenucleosidetransporter1pvent1forantimalarialdrugdevelopment AT frameij targetingtheplasmodiumvivaxequilibrativenucleosidetransporter1pvent1forantimalarialdrugdevelopment AT sosayvett targetingtheplasmodiumvivaxequilibrativenucleosidetransporter1pvent1forantimalarialdrugdevelopment AT akabasmylesh targetingtheplasmodiumvivaxequilibrativenucleosidetransporter1pvent1forantimalarialdrugdevelopment |