Effects of Single and Double Mutants in Human Glucose-6-Phosphate Dehydrogenase Variants Present in the Mexican Population: Biochemical and Structural Analysis

被引：15

作者：

Martinez-Rosas, Victor ^{[1
,2
]}

Valeria Juarez-Cruz, Merit ^{[3
]}

Jiovany Ramirez-Nava, Edson ^{[1
,4
]}

Hernandez-Ochoa, Beatriz ^{[2
,5
]}

Morales-Luna, Laura ^{[1
,4
]}

Gonzalez-Valdez, Abigail ^{[6
]}

Serrano-Posada, Hugo ^{[7
]}

Cardenas-Rodriguez, Noemi ^{[8
]}

Ortiz-Ramirez, Paulina ^{[1
]}

Centeno-Leija, Sara ^{[7
]}

Arreguin-Espinosa, Roberto ^{[9
]}

Cuevas-Cruz, Miguel ^{[9
]}

Ortega-Cuellar, Daniel ^{[10
]}

Perez de la Cruz, Veronica ^{[11
]}

Maria Rocha-Ramirez, Luz ^{[12
]}

Sierra-Palacios, Edgar ^{[13
]}

Angelica Castillo-Rodriguez, Rosa ^{[14
]}

Baeza-Ramirez, Isabel ^{[15
]}

Marcial-Quino, Jaime ^{[14
]}

Gomez-Manzo, Saul ^{[1
]}

机构：

[1] Inst Nacl Pediat, Secretaria Salud, Lab Bioquim Genet, Mexico City 04530, DF, Mexico

[2] Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Programa Posgrad Biomed & Biotecnol Mol, Mexico City 11340, DF, Mexico

[3] Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Posgrad Ciencias Alimentos, Dept Ingn Bioquim, Mexico City 11340, DF, Mexico

[4] Univ Nacl Autonoma Mexico, Posgrad Ciencias Biol, Mexico City 04510, DF, Mexico

[5] Secretaria Salud Mexico, Hosp Infantil Mexico Federico Gomez, Lab Inmunoquim, Mexico City 06720, DF, Mexico

[6] Univ Nacl Autonoma Mexico, Inst Invest Biomed, Dept Biol Mol & Biotecnol, Mexico City 04510, DF, Mexico

[7] Univ Colima, Tecnoparque CLQ, Lab Agrobiotecnol, Consejo Nacl Ciencia & Tecnol,CONACYT, Carretera Limones Loma de Juarez, Colima 28629, Mexico

[8] Inst Nacl Pediat, Secretaria Salud, Lab Neurociencias, Mexico City 04530, DF, Mexico

[9] Univ Nacl Autonoma Mexico, Inst Quim, Dept Quim Biomacromol, Mexico City 04510, DF, Mexico

[10] Inst Nacl Pediat, Secretaria Salud, Lab Nutr Expt, Mexico City 04530, DF, Mexico

[11] SSA, Inst Nacl Neurol & Neurocirug Manuel Velasco Suar, Dept Neuroquim, Mexico City 14269, DF, Mexico

[12] Hosp Infantil Mexico Dr Federico Gomez, Dept Infectol, Mexico City 06720, DF, Mexico

[13] Univ Autonoma Ciudad Mexico, Plantel Casa Libertad, Colegio Ciencias & Humanidades, Mexico City 09620, DF, Mexico

[14] Inst Nacl Pediat, Secretaria Salud, CONACYT, Consejo Nacl Ciencia & Tecnol, Mexico City 04530, DF, Mexico

[15] Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Lab Biomembranas, Mexico City 11340, DF, Mexico

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2020年 / 21卷 / 08期

关键词：

human G6PD mutants; G6PD deficiency; protein stability; structural characterization; catalytic activity; MUTATIONS; G6PD; STABILITY; DEFICIENCY; CLONING;

D O I：

10.3390/ijms21082732

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most frequent human enzymopathy, affecting over 400 million people globally. Worldwide, 217 mutations have been reported at the genetic level, and only 19 have been found in Mexico. The objective of this work was to contribute to the knowledge of the function and structure of three single natural variants (G6PD A+, G6PD San Luis Potosi, and G6PD Guadalajara) and a double mutant (G6PD Mount Sinai), each localized in a different region of the three-dimensional (3D) structure. In the functional characterization of the mutants, we observed a decrease in specific activity, protein expression and purification, catalytic efficiency, and substrate affinity in comparison with wild-type (WT) G6PD. Moreover, the analysis of the effect of all mutations on the structural stability showed that its presence increases denaturation and lability with temperature and it is more sensible to trypsin digestion protease and guanidine hydrochloride compared with WT G6PD. This could be explained by accelerated degradation of the variant enzymes due to reduced stability of the protein, as is shown in patients with G6PD deficiency.

引用

页数：23

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