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Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis
BACKGROUND: Nucleoside diphosphate kinase (NDK) is a housekeeping enzyme that plays key roles in nucleotide recycling and homeostasis in trypanosomatids. It is also secreted by the intracellular parasite Leishmania to modulate the host response. These functions make NDK an attractive target for drug...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4322457/ https://www.ncbi.nlm.nih.gov/pubmed/25643978 http://dx.doi.org/10.1186/s12900-015-0030-8 |
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author | Vieira, Plínio Salmazo de Giuseppe, Priscila Oliveira Murakami, Mario Tyago de Oliveira, Arthur Henrique Cavalcante |
author_facet | Vieira, Plínio Salmazo de Giuseppe, Priscila Oliveira Murakami, Mario Tyago de Oliveira, Arthur Henrique Cavalcante |
author_sort | Vieira, Plínio Salmazo |
collection | PubMed |
description | BACKGROUND: Nucleoside diphosphate kinase (NDK) is a housekeeping enzyme that plays key roles in nucleotide recycling and homeostasis in trypanosomatids. It is also secreted by the intracellular parasite Leishmania to modulate the host response. These functions make NDK an attractive target for drug design and for studies aiming at a better understanding of the mechanisms mediating host-pathogen interactions. RESULTS: We report the crystal structure and biophysical characterization of the NDK from Leishmania braziliensis (LbNDK). The subunit consists of six α-helices along with a core of four β-strands arranged in a β2β3β1β4 antiparallel topology order. In contrast to the NDK from L. major, the LbNDK C-terminal extension is partially unfolded. SAXS data showed that LbNDK forms hexamers in solution in the pH range from 7.0 to 4.0, a hydrodynamic behavior conserved in most eukaryotic NDKs. However, DSF assays show that acidification and alkalization decrease the hexamer stability. CONCLUSIONS: Our results support that LbNDK remains hexameric in pH conditions akin to that faced by this enzyme when secreted by Leishmania amastigotes in the parasitophorous vacuoles (pH 4.7 to 5.3). The unusual unfolded conformation of LbNDK C-terminus decreases the surface buried in the trimer interface exposing new regions that might be explored for the development of compounds designed to disturb enzyme oligomerization, which may impair the important nucleotide salvage pathway in these parasites. |
format | Online Article Text |
id | pubmed-4322457 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-43224572015-02-11 Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis Vieira, Plínio Salmazo de Giuseppe, Priscila Oliveira Murakami, Mario Tyago de Oliveira, Arthur Henrique Cavalcante BMC Struct Biol Research Article BACKGROUND: Nucleoside diphosphate kinase (NDK) is a housekeeping enzyme that plays key roles in nucleotide recycling and homeostasis in trypanosomatids. It is also secreted by the intracellular parasite Leishmania to modulate the host response. These functions make NDK an attractive target for drug design and for studies aiming at a better understanding of the mechanisms mediating host-pathogen interactions. RESULTS: We report the crystal structure and biophysical characterization of the NDK from Leishmania braziliensis (LbNDK). The subunit consists of six α-helices along with a core of four β-strands arranged in a β2β3β1β4 antiparallel topology order. In contrast to the NDK from L. major, the LbNDK C-terminal extension is partially unfolded. SAXS data showed that LbNDK forms hexamers in solution in the pH range from 7.0 to 4.0, a hydrodynamic behavior conserved in most eukaryotic NDKs. However, DSF assays show that acidification and alkalization decrease the hexamer stability. CONCLUSIONS: Our results support that LbNDK remains hexameric in pH conditions akin to that faced by this enzyme when secreted by Leishmania amastigotes in the parasitophorous vacuoles (pH 4.7 to 5.3). The unusual unfolded conformation of LbNDK C-terminus decreases the surface buried in the trimer interface exposing new regions that might be explored for the development of compounds designed to disturb enzyme oligomerization, which may impair the important nucleotide salvage pathway in these parasites. BioMed Central 2015-02-03 /pmc/articles/PMC4322457/ /pubmed/25643978 http://dx.doi.org/10.1186/s12900-015-0030-8 Text en © Vieira et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Vieira, Plínio Salmazo de Giuseppe, Priscila Oliveira Murakami, Mario Tyago de Oliveira, Arthur Henrique Cavalcante Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title | Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title_full | Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title_fullStr | Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title_full_unstemmed | Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title_short | Crystal structure and biophysical characterization of the nucleoside diphosphate kinase from Leishmania braziliensis |
title_sort | crystal structure and biophysical characterization of the nucleoside diphosphate kinase from leishmania braziliensis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4322457/ https://www.ncbi.nlm.nih.gov/pubmed/25643978 http://dx.doi.org/10.1186/s12900-015-0030-8 |
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