title: Mutations of Glucose-6-Phosphate Dehydrogenase Durham, Santa-Maria and A+ Variants Are Associated with Loss Functional and Structural Stability of the Protein.
abstract: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathy in the world. More than 160 mutations causing the disease have been identified, but only 10% of these variants have been studied at biochemical and biophysical levels. In this study we report on the functional and structural characterization of three naturally occurring variants corresponding to different classes of disease severity: Class I G6PD Durham, Class II G6PD Santa Maria, and Class III G6PD A+. The results showed that the G6PD Durham (severe deficiency), and the G6PD Santa Maria and A+ (less severe deficiency) (Class I, II and III, respectively) affect the catalytic efficiency of these enzymes, are more sensitive to temperature denaturing, and affect the stability of the overall protein when compared to the wild type WT-G6PD. In the variants, the exposure of more and buried hydrophobic pockets was induced and monitored with 8-Anilinonaphthalene-1-sulfonic acid (ANS) fluorescence, directly affecting the compaction of structure at different levels and probably reducing the stability of the protein. The degree of functional and structural perturbation by each variant correlates with the clinical severity reported in different patients.
authors.affiliation: Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. saulmanzo@ciencias.unam.mx.
authors.forename: Saúl
authors.initials: S
authors.lastname: Gómez-Manzo
authors.affiliation: CONACYT Research Fellow-Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. jmarcialqu@conacyt.mx.
authors.forename: Jaime
authors.initials: J
authors.lastname: Marcial-Quino
authors.affiliation: Laboratorio de Neurociencias, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. america_vc@yahoo.com.mx.
authors.forename: America
authors.initials: A
authors.lastname: Vanoye-Carlo
authors.affiliation: Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. sergioenr@gmail.com.
authors.forename: Sergio
authors.initials: S
authors.lastname: Enríquez-Flores
authors.affiliation: Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. ignaciodelamora@yahoo.com.mx.
authors.forename: Ignacio
authors.initials: I
authors.lastname: De la Mora-De la Mora
authors.affiliation: Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de Mexico, Mexico D.F. 04510, Mexico. abigaila@correo.biomedicas.unam.mx.
authors.forename: Abigail
authors.initials: A
authors.lastname: González-Valdez
authors.affiliation: Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. itzheltorres@hotmail.com.
authors.forename: Itzhel
authors.initials: I
authors.lastname: García-Torres
authors.affiliation: Laboratorio de Bioquímica Genética, Instituto Nacional de Pediatría, Mexico D.F. 04530, Mexico. razer_45r@hotmail.com.
authors.forename: Víctor
authors.initials: V
authors.lastname: Martínez-Rosas
authors.affiliation: Colegio de Ciencias y Humanidades, Plantel Casa Libertad, UACM, Mexico D.F.09620, Mexico. edgar.sierra@uacm.edu.mx.
authors.forename: Edgar
authors.initials: E
authors.lastname: Sierra-Palacios
authors.affiliation: Departamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de Mexico, México, D.F. 04510, Mexico. ferlaz@hotmail.com.
authors.forename: Fernando
authors.initials: F
authors.lastname: Lazcano-Pérez
authors.affiliation: Departamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de Mexico, México, D.F. 04510, Mexico. e-rodriguez-bustamante@ciencias.unam.mx.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Rodríguez-Bustamante
authors.affiliation: Departamento de Química de Biomacromoléculas, Instituto de Química, Universidad Nacional Autónoma de Mexico, México, D.F. 04510, Mexico. arrespin@unam.mx.
authors.forename: Roberto
authors.initials: R
authors.lastname: Arreguin-Espinosa
mesh_terms: D002384:Catalysis; D004789:Enzyme Activation; D015870:Gene Expression; D014644:Genetic Variation; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D055550:Protein Stability; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D013816:Thermodynamics
chemical_list: D011994:Recombinant Proteins; D005954:Glucosephosphate Dehydrogenase
keywords: G6PD deficiency; G6PD-variants; kinetic-properties; recombinant expression; stability