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Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP

Helicobacter pylori (H. pylori) has been proposed as the foremost risk factor for the development of gastric cancer. We found that H. pylori express the enzyme glucose-6-phosphate dehydrogenase (HpG6PD), which participates in glucose metabolism via the pentose phosphate pathway. Thus, we hypothesize...

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Autores principales: Ortiz-Ramírez, Paulina, Hernández-Ochoa, Beatriz, Ortega-Cuellar, Daniel, González-Valdez, Abigail, Martínez-Rosas, Víctor, Morales-Luna, Laura, Arreguin-Espinosa, Roberto, Castillo-Rodríguez, Rosa Angélica, Canseco-Ávila, Luis Miguel, Cárdenas-Rodríguez, Noemi, Pérez de la Cruz, Verónica, Montiel-González, Alba Mónica, Gómez-Chávez, Fernando, Gómez-Manzo, Saúl
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9323780/
https://www.ncbi.nlm.nih.gov/pubmed/35889079
http://dx.doi.org/10.3390/microorganisms10071359
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author Ortiz-Ramírez, Paulina
Hernández-Ochoa, Beatriz
Ortega-Cuellar, Daniel
González-Valdez, Abigail
Martínez-Rosas, Víctor
Morales-Luna, Laura
Arreguin-Espinosa, Roberto
Castillo-Rodríguez, Rosa Angélica
Canseco-Ávila, Luis Miguel
Cárdenas-Rodríguez, Noemi
Pérez de la Cruz, Verónica
Montiel-González, Alba Mónica
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
author_facet Ortiz-Ramírez, Paulina
Hernández-Ochoa, Beatriz
Ortega-Cuellar, Daniel
González-Valdez, Abigail
Martínez-Rosas, Víctor
Morales-Luna, Laura
Arreguin-Espinosa, Roberto
Castillo-Rodríguez, Rosa Angélica
Canseco-Ávila, Luis Miguel
Cárdenas-Rodríguez, Noemi
Pérez de la Cruz, Verónica
Montiel-González, Alba Mónica
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
author_sort Ortiz-Ramírez, Paulina
collection PubMed
description Helicobacter pylori (H. pylori) has been proposed as the foremost risk factor for the development of gastric cancer. We found that H. pylori express the enzyme glucose-6-phosphate dehydrogenase (HpG6PD), which participates in glucose metabolism via the pentose phosphate pathway. Thus, we hypothesized that if the biochemical and physicochemical characteristics of HpG6PD contrast with the host G6PD (human G6PD, HsG6PD), HpG6PD becomes a potential target for novel drugs against H. pylori. In this work, we characterized the biochemical properties of the HpG6PD from the H. pylori strain 29CaP and expressed the active recombinant protein, to analyze its steady-state kinetics, thermostability, and biophysical aspects. In addition, we analyzed the HpG6PD in silico structural properties to compare them with those of the HsG6PD. The optimal pH for enzyme activity was 7.5, with a T(1/2) of 46.6 °C, at an optimum stability temperature of 37 °C. The apparent K(m) values calculated for G6P and NADP(+) were 75.0 and 12.8 µM, respectively. G6P does not protect HpG6PD from trypsin digestion, but NADP(+) does protect the enzyme from trypsin and guanidine hydrochloride (Gdn-HCl). The biochemical characterization of HpG6PD contributes to knowledge regarding H. pylori metabolism and opens up the possibility of using this enzyme as a potential target for specific and efficient treatment against this pathogen; structural alignment indicates that the three-dimensional (3D) homodimer model of the G6PD protein from H. pylori is different from the 3D G6PD of Homo sapiens.
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spelling pubmed-93237802022-07-27 Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP Ortiz-Ramírez, Paulina Hernández-Ochoa, Beatriz Ortega-Cuellar, Daniel González-Valdez, Abigail Martínez-Rosas, Víctor Morales-Luna, Laura Arreguin-Espinosa, Roberto Castillo-Rodríguez, Rosa Angélica Canseco-Ávila, Luis Miguel Cárdenas-Rodríguez, Noemi Pérez de la Cruz, Verónica Montiel-González, Alba Mónica Gómez-Chávez, Fernando Gómez-Manzo, Saúl Microorganisms Article Helicobacter pylori (H. pylori) has been proposed as the foremost risk factor for the development of gastric cancer. We found that H. pylori express the enzyme glucose-6-phosphate dehydrogenase (HpG6PD), which participates in glucose metabolism via the pentose phosphate pathway. Thus, we hypothesized that if the biochemical and physicochemical characteristics of HpG6PD contrast with the host G6PD (human G6PD, HsG6PD), HpG6PD becomes a potential target for novel drugs against H. pylori. In this work, we characterized the biochemical properties of the HpG6PD from the H. pylori strain 29CaP and expressed the active recombinant protein, to analyze its steady-state kinetics, thermostability, and biophysical aspects. In addition, we analyzed the HpG6PD in silico structural properties to compare them with those of the HsG6PD. The optimal pH for enzyme activity was 7.5, with a T(1/2) of 46.6 °C, at an optimum stability temperature of 37 °C. The apparent K(m) values calculated for G6P and NADP(+) were 75.0 and 12.8 µM, respectively. G6P does not protect HpG6PD from trypsin digestion, but NADP(+) does protect the enzyme from trypsin and guanidine hydrochloride (Gdn-HCl). The biochemical characterization of HpG6PD contributes to knowledge regarding H. pylori metabolism and opens up the possibility of using this enzyme as a potential target for specific and efficient treatment against this pathogen; structural alignment indicates that the three-dimensional (3D) homodimer model of the G6PD protein from H. pylori is different from the 3D G6PD of Homo sapiens. MDPI 2022-07-06 /pmc/articles/PMC9323780/ /pubmed/35889079 http://dx.doi.org/10.3390/microorganisms10071359 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ortiz-Ramírez, Paulina
Hernández-Ochoa, Beatriz
Ortega-Cuellar, Daniel
González-Valdez, Abigail
Martínez-Rosas, Víctor
Morales-Luna, Laura
Arreguin-Espinosa, Roberto
Castillo-Rodríguez, Rosa Angélica
Canseco-Ávila, Luis Miguel
Cárdenas-Rodríguez, Noemi
Pérez de la Cruz, Verónica
Montiel-González, Alba Mónica
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title_full Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title_fullStr Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title_full_unstemmed Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title_short Biochemical and Kinetic Characterization of the Glucose-6-Phosphate Dehydrogenase from Helicobacter pylori Strain 29CaP
title_sort biochemical and kinetic characterization of the glucose-6-phosphate dehydrogenase from helicobacter pylori strain 29cap
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9323780/
https://www.ncbi.nlm.nih.gov/pubmed/35889079
http://dx.doi.org/10.3390/microorganisms10071359
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