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
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...
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Published in | Applied biochemistry and biotechnology Vol. 182; no. 4; pp. 1478 - 1490 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
New York
Springer US
01.08.2017
Springer Nature B.V |
Subjects | |
Online Access | Get full text |
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Summary: | 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
k
m
and
V
max
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 IC
50
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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0273-2289 1559-0291 |
DOI: | 10.1007/s12010-017-2412-9 |