Internet Archive Scholar

Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia

Laura Morales-Luna, Hugo Serrano-Posada, Abigail González-Valdez, Daniel Ortega-Cuellar, America Vanoye-Carlo, Beatriz Hernández-Ochoa, Edgar Sierra-Palacios, Yadira Rufino-González, Rosa Castillo-Rodríguez, Verónica Pérez de la Cruz, Liliana Moreno-Vargas, Diego Prada-Gracia (+2 others)
2018 International Journal of Molecular Sciences  

Preserved Fulltext

fulltext thumbnail
Web Archive Capture PDF (3.0 MB)
https://web.archive.org/web/20191019215009/http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC6165198&blobtype=pdf

A copy of this work was available on the public web and has been preserved in the Wayback Machine. The capture dates from 2019; you can also visit the original URL. The file type is application/pdf.

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
more » ... 6PD::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.
doi:10.3390/ijms19092518 pmid:30149622 fatcat:4bc4aeblxfbnbknjfhxminivu4
DOAJ
Citation

Laura Morales-Luna, Hugo Serrano-Posada, Abigail González-Valdez, Daniel Ortega-Cuellar, America Vanoye-Carlo, Beatriz Hernández-Ochoa, Edgar Sierra-Palacios, Yadira Rufino-González, Rosa Castillo-Rodríguez, Verónica Pérez de la Cruz, Liliana Moreno-Vargas, Diego Prada-Gracia, Jaime Marcial-Quino, Saúl Gómez-Manzo. "Biochemical Characterization and Structural Modeling of Fused Glucose-6-Phosphate Dehydrogenase-Phosphogluconolactonase from Giardia lamblia." International Journal of Molecular Sciences 19.9 (2018) 2518