Functional and Biochemical Characterization of Three Recombinant Human Glucose-6-Phosphate Dehydrogenase Mutants: Zacatecas, Vanua-Lava and Viangchan.

Abstract:

:Glucose-6-phosphate dehydrogenase (G6PD) deficiency in humans causes severe disease, varying from mostly asymptomatic individuals to patients showing neonatal jaundice, acute hemolysis episodes or chronic nonspherocytic hemolytic anemia. In order to understand the effect of the mutations in G6PD gene function and its relation with G6PD deficiency severity, we report the construction, cloning and expression as well as the detailed kinetic and stability characterization of three purified clinical variants of G6PD that present in the Mexican population: G6PD Zacatecas (Class I), Vanua-Lava (Class II) and Viangchan (Class II). For all the G6PD mutants, we obtained low purification yield and altered kinetic parameters compared with Wild Type (WT). Our results show that the mutations, regardless of the distance from the active site where they are located, affect the catalytic properties and structural parameters and that these changes could be associated with the clinical presentation of the deficiency. Specifically, the structural characterization of the G6PD Zacatecas mutant suggests that the R257L mutation have a strong effect on the global stability of G6PD favoring an unstable active site. Using computational analysis, we offer a molecular explanation of the effects of these mutations on the active site.

journal_name

Int J Mol Sci

authors

Gómez-Manzo S,Marcial-Quino J,Vanoye-Carlo A,Serrano-Posada H,González-Valdez A,Martínez-Rosas V,Hernández-Ochoa B,Sierra-Palacios E,Castillo-Rodríguez RA,Cuevas-Cruz M,Rodríguez-Bustamante E,Arreguin-Espinosa R

doi

10.3390/ijms17050787

subject

Has Abstract

pub_date

2016-05-21 00:00:00

issue

5

issn

1422-0067

pii

ijms17050787

journal_volume

17

pub_type

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