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Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme

Giardia lamblia is a single-celled eukaryotic parasite with a small genome and is considered an early divergent eukaryote. The pentose phosphate pathway (PPP) plays an essential role in the oxidative stress defense of the parasite and the production of ribose-5-phosphate. In this parasite, the gluco...

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Autores principales: Morales-Luna, Laura, González-Valdez, Abigail, Hernández-Ochoa, Beatriz, Arreguin-Espinosa, Roberto, Ortega-Cuellar, Daniel, Castillo-Rodríguez, Rosa Angélica, Martínez-Rosas, Víctor, Cárdenas-Rodríguez, Noemi, Enríquez-Flores, Sergio, Canseco-Ávila, Luis Miguel, de la Cruz, Verónica Pérez, Gómez-Chávez, Fernando, Gómez-Manzo, Saúl
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399836/
https://www.ncbi.nlm.nih.gov/pubmed/34442758
http://dx.doi.org/10.3390/microorganisms9081678
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author Morales-Luna, Laura
González-Valdez, Abigail
Hernández-Ochoa, Beatriz
Arreguin-Espinosa, Roberto
Ortega-Cuellar, Daniel
Castillo-Rodríguez, Rosa Angélica
Martínez-Rosas, Víctor
Cárdenas-Rodríguez, Noemi
Enríquez-Flores, Sergio
Canseco-Ávila, Luis Miguel
de la Cruz, Verónica Pérez
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
author_facet Morales-Luna, Laura
González-Valdez, Abigail
Hernández-Ochoa, Beatriz
Arreguin-Espinosa, Roberto
Ortega-Cuellar, Daniel
Castillo-Rodríguez, Rosa Angélica
Martínez-Rosas, Víctor
Cárdenas-Rodríguez, Noemi
Enríquez-Flores, Sergio
Canseco-Ávila, Luis Miguel
de la Cruz, Verónica Pérez
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
author_sort Morales-Luna, Laura
collection PubMed
description Giardia lamblia is a single-celled eukaryotic parasite with a small genome and is considered an early divergent eukaryote. The pentose phosphate pathway (PPP) plays an essential role in the oxidative stress defense of the parasite and the production of ribose-5-phosphate. In this parasite, the glucose-6-phosphate dehydrogenase (G6PD) is fused with the 6-phosphogluconolactonase (6PGL) enzyme, generating the enzyme named G6PD::6PGL that catalyzes the first two steps of the PPP. Here, we report that the G6PD::6PGL is a bifunctional enzyme with two catalytically active sites. We performed the kinetic characterization of both domains in the fused G6PD::6PGL enzyme, as well as the individual cloned G6PD. The results suggest that the catalytic activity of G6PD and 6PGL domains in the G6PD::6PGL enzyme are more efficient than the individual proteins. Additionally, using enzymatic and mass spectrometry assays, we found that the final metabolites of the catalytic reaction of the G6PD::6PGL are 6-phosphoglucono-δ-lactone and 6-phosphogluconate. Finally, we propose the reaction mechanism in which the G6PD domain performs the catalysis, releasing 6-phosphoglucono-δ-lactone to the reaction medium. Then, this metabolite binds to the 6PGL domain catalyzing the hydrolysis reaction and generating 6-phosphogluconate. The structural difference between the G. lamblia fused enzyme G6PD::6PGL with the human G6PD indicate that the G6PD::6PGL is a potential drug target for the rational synthesis of novels anti-Giardia drugs.
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spelling pubmed-83998362021-08-29 Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme Morales-Luna, Laura González-Valdez, Abigail Hernández-Ochoa, Beatriz Arreguin-Espinosa, Roberto Ortega-Cuellar, Daniel Castillo-Rodríguez, Rosa Angélica Martínez-Rosas, Víctor Cárdenas-Rodríguez, Noemi Enríquez-Flores, Sergio Canseco-Ávila, Luis Miguel de la Cruz, Verónica Pérez Gómez-Chávez, Fernando Gómez-Manzo, Saúl Microorganisms Article Giardia lamblia is a single-celled eukaryotic parasite with a small genome and is considered an early divergent eukaryote. The pentose phosphate pathway (PPP) plays an essential role in the oxidative stress defense of the parasite and the production of ribose-5-phosphate. In this parasite, the glucose-6-phosphate dehydrogenase (G6PD) is fused with the 6-phosphogluconolactonase (6PGL) enzyme, generating the enzyme named G6PD::6PGL that catalyzes the first two steps of the PPP. Here, we report that the G6PD::6PGL is a bifunctional enzyme with two catalytically active sites. We performed the kinetic characterization of both domains in the fused G6PD::6PGL enzyme, as well as the individual cloned G6PD. The results suggest that the catalytic activity of G6PD and 6PGL domains in the G6PD::6PGL enzyme are more efficient than the individual proteins. Additionally, using enzymatic and mass spectrometry assays, we found that the final metabolites of the catalytic reaction of the G6PD::6PGL are 6-phosphoglucono-δ-lactone and 6-phosphogluconate. Finally, we propose the reaction mechanism in which the G6PD domain performs the catalysis, releasing 6-phosphoglucono-δ-lactone to the reaction medium. Then, this metabolite binds to the 6PGL domain catalyzing the hydrolysis reaction and generating 6-phosphogluconate. The structural difference between the G. lamblia fused enzyme G6PD::6PGL with the human G6PD indicate that the G6PD::6PGL is a potential drug target for the rational synthesis of novels anti-Giardia drugs. MDPI 2021-08-07 /pmc/articles/PMC8399836/ /pubmed/34442758 http://dx.doi.org/10.3390/microorganisms9081678 Text en © 2021 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
Morales-Luna, Laura
González-Valdez, Abigail
Hernández-Ochoa, Beatriz
Arreguin-Espinosa, Roberto
Ortega-Cuellar, Daniel
Castillo-Rodríguez, Rosa Angélica
Martínez-Rosas, Víctor
Cárdenas-Rodríguez, Noemi
Enríquez-Flores, Sergio
Canseco-Ávila, Luis Miguel
de la Cruz, Verónica Pérez
Gómez-Chávez, Fernando
Gómez-Manzo, Saúl
Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title_full Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title_fullStr Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title_full_unstemmed Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title_short Glucose-6-Phosphate Dehydrogenase::6-Phosphogluconolactonase from the Parasite Giardia lamblia. A Molecular and Biochemical Perspective of a Fused Enzyme
title_sort glucose-6-phosphate dehydrogenase::6-phosphogluconolactonase from the parasite giardia lamblia. a molecular and biochemical perspective of a fused enzyme
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399836/
https://www.ncbi.nlm.nih.gov/pubmed/34442758
http://dx.doi.org/10.3390/microorganisms9081678
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