TY - JOUR
T1 - 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
AU - Andrade-Pavón, Dulce
AU - Cuevas-Hernández, Roberto I.
AU - Trujillo-Ferrara, José G.
AU - Hernández-Rodríguez, César
AU - Ibarra, J. Antonio
AU - Villa-Tanaca, Lourdes
N1 - Publisher Copyright:
© 2017, Springer Science+Business Media New York.
PY - 2017/8/1
Y1 - 2017/8/1
N2 - 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 km and Vmax for the HMG-CoA were 6.5 μM and 2.26 × 10−3 μM min−1, respectively. Recombinant CgHMGR was inhibited by simvastatin presenting an IC50 at 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.
AB - 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 km and Vmax for the HMG-CoA were 6.5 μM and 2.26 × 10−3 μM min−1, respectively. Recombinant CgHMGR was inhibited by simvastatin presenting an IC50 at 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.
KW - Antifungal
KW - Candida glabrata
KW - EC 1.1.1.34
KW - HMGR inhibitors
KW - Heterologous expression
KW - Recombinant HMGR
UR - http://www.scopus.com/inward/record.url?scp=85010972101&partnerID=8YFLogxK
U2 - 10.1007/s12010-017-2412-9
DO - 10.1007/s12010-017-2412-9
M3 - Artículo
C2 - 28138930
AN - SCOPUS:85010972101
SN - 0273-2289
VL - 182
SP - 1478
EP - 1490
JO - Applied Biochemistry and Biotechnology
JF - Applied Biochemistry and Biotechnology
IS - 4
ER -