Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major.
Deoxyhypusine synthase (DHS) catalyzes the first step of the post-translational modification of eukaryotic translation factor 5A (eIF5A), which is the only known protein containing the amino acid hypusine. Both proteins are essential for eukaryotic cell viability, and DHS has been suggested as a goo...
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ftdoajarticles:oai:doaj.org/article:d575f849eb914816adf7653b1a595e86 2023-05-15T15:12:23+02:00 Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. Suélen Fernandes Silva Angélica Hollunder Klippel Priscila Zonzini Ramos André da Silva Santiago Sandro Roberto Valentini Mario Henrique Bengtson Katlin Brauer Massirer Elizabeth Bilsland Rafael Miguez Couñago Cleslei Fernando Zanelli 2020-10-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0008762 https://doaj.org/article/d575f849eb914816adf7653b1a595e86 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0008762 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008762 https://doaj.org/article/d575f849eb914816adf7653b1a595e86 PLoS Neglected Tropical Diseases, Vol 14, Iss 10, p e0008762 (2020) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2020 ftdoajarticles https://doi.org/10.1371/journal.pntd.0008762 2022-12-31T05:52:29Z Deoxyhypusine synthase (DHS) catalyzes the first step of the post-translational modification of eukaryotic translation factor 5A (eIF5A), which is the only known protein containing the amino acid hypusine. Both proteins are essential for eukaryotic cell viability, and DHS has been suggested as a good candidate target for small molecule-based therapies against eukaryotic pathogens. In this work, we focused on the DHS enzymes from Brugia malayi and Leishmania major, the causative agents of lymphatic filariasis and cutaneous leishmaniasis, respectively. To enable B. malayi (Bm)DHS for future target-based drug discovery programs, we determined its crystal structure bound to cofactor NAD+. We also reported an in vitro biochemical assay for this enzyme that is amenable to a high-throughput screening format. The L. major genome encodes two DHS paralogs, and attempts to produce them recombinantly in bacterial cells were not successful. Nevertheless, we showed that ectopic expression of both LmDHS paralogs can rescue yeast cells lacking the endogenous DHS-encoding gene (dys1). Thus, functionally complemented dys1Δ yeast mutants can be used to screen for new inhibitors of the L. major enzyme. We used the known human DHS inhibitor GC7 to validate both in vitro and yeast-based DHS assays. Our results show that BmDHS is a homotetrameric enzyme that shares many features with its human homologue, whereas LmDHS paralogs are likely to form a heterotetrameric complex and have a distinct regulatory mechanism. We expect our work to facilitate the identification and development of new DHS inhibitors that can be used to validate these enzymes as vulnerable targets for therapeutic interventions against B. malayi and L. major infections. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 14 10 e0008762 |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Suélen Fernandes Silva Angélica Hollunder Klippel Priscila Zonzini Ramos André da Silva Santiago Sandro Roberto Valentini Mario Henrique Bengtson Katlin Brauer Massirer Elizabeth Bilsland Rafael Miguez Couñago Cleslei Fernando Zanelli Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Deoxyhypusine synthase (DHS) catalyzes the first step of the post-translational modification of eukaryotic translation factor 5A (eIF5A), which is the only known protein containing the amino acid hypusine. Both proteins are essential for eukaryotic cell viability, and DHS has been suggested as a good candidate target for small molecule-based therapies against eukaryotic pathogens. In this work, we focused on the DHS enzymes from Brugia malayi and Leishmania major, the causative agents of lymphatic filariasis and cutaneous leishmaniasis, respectively. To enable B. malayi (Bm)DHS for future target-based drug discovery programs, we determined its crystal structure bound to cofactor NAD+. We also reported an in vitro biochemical assay for this enzyme that is amenable to a high-throughput screening format. The L. major genome encodes two DHS paralogs, and attempts to produce them recombinantly in bacterial cells were not successful. Nevertheless, we showed that ectopic expression of both LmDHS paralogs can rescue yeast cells lacking the endogenous DHS-encoding gene (dys1). Thus, functionally complemented dys1Δ yeast mutants can be used to screen for new inhibitors of the L. major enzyme. We used the known human DHS inhibitor GC7 to validate both in vitro and yeast-based DHS assays. Our results show that BmDHS is a homotetrameric enzyme that shares many features with its human homologue, whereas LmDHS paralogs are likely to form a heterotetrameric complex and have a distinct regulatory mechanism. We expect our work to facilitate the identification and development of new DHS inhibitors that can be used to validate these enzymes as vulnerable targets for therapeutic interventions against B. malayi and L. major infections. |
format |
Article in Journal/Newspaper |
author |
Suélen Fernandes Silva Angélica Hollunder Klippel Priscila Zonzini Ramos André da Silva Santiago Sandro Roberto Valentini Mario Henrique Bengtson Katlin Brauer Massirer Elizabeth Bilsland Rafael Miguez Couñago Cleslei Fernando Zanelli |
author_facet |
Suélen Fernandes Silva Angélica Hollunder Klippel Priscila Zonzini Ramos André da Silva Santiago Sandro Roberto Valentini Mario Henrique Bengtson Katlin Brauer Massirer Elizabeth Bilsland Rafael Miguez Couñago Cleslei Fernando Zanelli |
author_sort |
Suélen Fernandes Silva |
title |
Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
title_short |
Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
title_full |
Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
title_fullStr |
Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
title_full_unstemmed |
Structural features and development of an assay platform of the parasite target deoxyhypusine synthase of Brugia malayi and Leishmania major. |
title_sort |
structural features and development of an assay platform of the parasite target deoxyhypusine synthase of brugia malayi and leishmania major. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2020 |
url |
https://doi.org/10.1371/journal.pntd.0008762 https://doaj.org/article/d575f849eb914816adf7653b1a595e86 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 14, Iss 10, p e0008762 (2020) |
op_relation |
https://doi.org/10.1371/journal.pntd.0008762 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0008762 https://doaj.org/article/d575f849eb914816adf7653b1a595e86 |
op_doi |
https://doi.org/10.1371/journal.pntd.0008762 |
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PLOS Neglected Tropical Diseases |
container_volume |
14 |
container_issue |
10 |
container_start_page |
e0008762 |
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1766343076560764928 |