Título

Biochemical characterization and structural modeling of fused glucose-6-phosphate dehydrogenase-phosphogluconolactonase from giardia lamblia

Autor

Laura Morales-Luna

HUGO JAVIER SERRANO POSADA

ALEJANDRA ABIGAIL GONZALEZ VALDEZ

DANIEL DAGOBERTO ORTEGA CUELLAR

AMERICA VANOYE CARLO

BEATRIZ HERNANDEZ OCHOA

EDGAR DEL CARMEN SIERRA PALACIOS

YADIRA RUFINO GONZALEZ

ROSA ANGELICA CASTILLO RODRIGUEZ

VERONICA PEREZ DE LA CRUZ

LILIANA MARISOL MORENO VARGAS

DIEGO PRADA GRACIA

JAIME MARCIAL QUINO

SAUL GOMEZ MANZO

Nivel de Acceso

Acceso Abierto

Licencia

http://creativecommons.org/licenses/by/4.0

Materias

BIOQUÍMICA - (CTI) Giardia lamblia - (LCHS) G6PD - (SIN FUENTE) Purification - (SIN FUENTE) Recombinant expression - (SIN FUENTE) Bioinformatics analysis - (SIN FUENTE) Three-dimensional structure - (SIN FUENTE)

Resumen o descripción

Glucose-6-phosphate dehydrogenase (G6PD) is the first enzyme in the pentose phosphate pathway and is highly relevant in the metabolism of Giardia lamblia. Previous reports suggested that the G6PD gene is fused with the 6-phosphogluconolactonase (6PGL) gene (6pgl). Therefore, in this work, we decided to characterize the fused G6PD-6PGL protein in Giardia lamblia. First, the gene of g6pd fused with the 6pgl gene (6gpd::6pgl) was isolated from trophozoites of Giardia lamblia and the corresponding G6PD::6PGL protein was overexpressed and purified in Escherichia coli. Then, we characterized the native oligomeric state of the G6PD::6PGL protein in solution and we found a catalytic dimer with an optimum pH of 8.75. Furthermore, we determined the steady-state kinetic parameters for the G6PD domain and measured the thermal stability of the protein in both the presence and absence of guanidine hydrochloride (Gdn-HCl) and observed that the G6PD::6PGL protein showed alterations in the stability, secondary structure, and tertiary structure in the presence of Gdn-HCl. Finally, computer modeling studies revealed unique structural and functional features, which clearly established the differences between G6PD::6PGL protein from G. lamblia and the human G6PD enzyme, proving that the model can be used for the design of new drugs with antigiardiasic activity. These results broaden the perspective for future studies of the function of the protein and its effect on the metabolism of this parasite as a potential pharmacological target.

Editor

MDPI

Fecha de publicación

25 de agosto de 2018

Tipo de publicación

Artículo

Recurso de información

http://biblos.ucol.mx//handle/123456789/81

Idioma

Inglés

Repositorio Orígen

Repositorio institucional de la Universidad de Colima

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