title: Recombinant 3-Hydroxy 3-Methyl Glutaryl-CoA Reductase from Candida glabrata (Rec-CgHMGR) Obtained by Heterologous Expression, as a Novel Therapeutic Target Model for Testing Synthetic Drugs.
abstract: The enzyme 3-hydroxy-3-methyl-glutaryl CoA reductase (HMGR) is a glycoprotein of the endoplasmic reticulum that participates in the mevalonate pathway, the precursor of cholesterol in human and ergosterol in fungi. This enzyme has three domains: transmembrane, binding, and soluble. In this study, we expressed and purified the soluble fraction of the HMGR enzyme from Candida glabrata (CgHMGR) in an Escherichia coli heterologous system and used it as a model for studying its inhibitory activity. The soluble fraction of CgHMGR was fused to the maltose binding protein (MBP), purified, and characterized. Optimal pH was 8.0, and its optimal temperature activity was 37 °C. The kmand Vmaxfor the HMG-CoA were 6.5 μM and 2.26 × 10-3μM min-1, respectively. Recombinant CgHMGR was inhibited by simvastatin presenting an IC50at 14.5 μM. In conclusion, our findings suggest that the recombinant HMGR version from C. glabrata may be used as a study model system for HMGR inhibitors such as statins and newly synthesized inhibitor compounds that might be used in the treatment of hypercholesterolemia or mycosis.
authors.affiliation: Laboratorio de Genética Microbiana, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. de Carpio y Plan de Ayala. Col. Sto. Tomás, 11340, Ciudad de México, DF, Mexico.
authors.forename: Dulce
authors.initials: D
authors.lastname: Andrade-Pavón
authors.affiliation: Laboratorio de Investigación en Bioquímica, Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón, Casco de Santo Tomás, 11340, Ciudad de México, Mexico.
authors.forename: Roberto I
authors.initials: RI
authors.lastname: Cuevas-Hernández
authors.affiliation: Laboratorio de Investigación en Bioquímica, Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Instituto Politécnico Nacional, Plan de San Luis y Díaz Mirón, Casco de Santo Tomás, 11340, Ciudad de México, Mexico.
authors.forename: José G
authors.initials: JG
authors.lastname: Trujillo-Ferrara
authors.affiliation: Laboratorio de Genética Microbiana, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. de Carpio y Plan de Ayala. Col. Sto. Tomás, 11340, Ciudad de México, DF, Mexico.
authors.forename: César
authors.initials: C
authors.lastname: Hernández-Rodríguez
authors.affiliation: Laboratorio de Genética Microbiana, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. de Carpio y Plan de Ayala. Col. Sto. Tomás, 11340, Ciudad de México, DF, Mexico. jaig19@gmail.com.
authors.forename: J Antonio
authors.initials: JA
authors.lastname: Ibarra
authors.affiliation: Laboratorio de Genética Microbiana, Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prol. de Carpio y Plan de Ayala. Col. Sto. Tomás, 11340, Ciudad de México, DF, Mexico. mvillat@ipn.mx.
authors.forename: Lourdes
authors.identifier: http://orcid.org/0000-0003-0055-2991
authors.initials: L
authors.lastname: Villa-Tanaca
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D041221:Candida glabrata / Q000201:enzymology* / Q000235:genetics; D004353:Drug Evaluation, Preclinical; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006903:Hydroxymethylglutaryl CoA Reductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D007700:Kinetics; D058619:Maltose-Binding Proteins / Q000378:metabolism; D008958:Models, Molecular; D058990:Molecular Targeted Therapy; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D019821:Simvastatin / Q000494:pharmacology; D012995:Solubility; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D019821:Simvastatin; D006903:Hydroxymethylglutaryl CoA Reductases
keywords: Antifungal; Candida glabrata; EC 1.1.1.34; HMGR inhibitors; Heterologous expression; Recombinant HMGR