Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Vieira, Plínio Salmazo, de Giuseppe, Priscila Oliveira, Murakami, Mario Tyago, de Oliveira, Arthur Henrique Cavalcante
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
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
_version_ 1782356384411549696
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
work_keys_str_mv AT vieirapliniosalmazo crystalstructureandbiophysicalcharacterizationofthenucleosidediphosphatekinasefromleishmaniabraziliensis
AT degiuseppepriscilaoliveira crystalstructureandbiophysicalcharacterizationofthenucleosidediphosphatekinasefromleishmaniabraziliensis
AT murakamimariotyago crystalstructureandbiophysicalcharacterizationofthenucleosidediphosphatekinasefromleishmaniabraziliensis
AT deoliveiraarthurhenriquecavalcante crystalstructureandbiophysicalcharacterizationofthenucleosidediphosphatekinasefromleishmaniabraziliensis