Inhibition of recombinant enzyme 3-hydroxy-3-methylglutaryl-CoA reductase from Candida glabrata by α-asarone-based synthetic compounds as antifungal agents

Andrade-Pavón, Dulce; Ortiz‐Álvarez, Jossue; Sánchez-Sandoval, Eugenia; Tamariz, Joaquı́n; Hernández, César Arranz; Ibarra, J. Antonio; Villa‐Tanaca, Lourdes

doi:10.1016/j.jbiotec.2019.01.008

Cited by 15 publications

(38 citation statements)

References 28 publications

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“…In its wild type form, HMGRCg can catalyze HMG-CoA to mevalonate through the oxidation of NADPH. 10,11 The structures of the mutated proteins were validated by stereo-chemical restriction determinations on Ramachandran plots, thus providing an a priori approximation to the 3D structure of the corresponding peptides, a necessary step for their in-silico analysis. 11 Statins and alpha-asarone are competitive inhibitors of HMGRs, blocking access to the HMG-CoA substrate.…”

Section: Discussionmentioning

confidence: 99%

“…10,11 The structures of the mutated proteins were validated by stereo-chemical restriction determinations on Ramachandran plots, thus providing an a priori approximation to the 3D structure of the corresponding peptides, a necessary step for their in-silico analysis. 11 Statins and alpha-asarone are competitive inhibitors of HMGRs, blocking access to the HMG-CoA substrate. 1,3,10,11 Simvastatin (like other statins) and alpha-asarone have an HMG-like moiety capable of replacing the thioester oxygen atom found in the HMG-CoA substrate.…”

Section: Discussionmentioning

confidence: 99%

“…11 Statins and alpha-asarone are competitive inhibitors of HMGRs, blocking access to the HMG-CoA substrate. 1,3,10,11 Simvastatin (like other statins) and alpha-asarone have an HMG-like moiety capable of replacing the thioester oxygen atom found in the HMG-CoA substrate. 3 The HMGRCg mutant in the substrate binding motif, E711Q, displayed the lowest binding energies in relation to the ligands presently examined: simvastatin, alpha-asarone and the substrate HMG-CoA.…”

Section: Discussionmentioning

confidence: 99%

“…The reduced enzymatic activity and binding energy of this mutant was not unexpected, considering that the three ligands currently tested interact with the EGCLVAS motif (of the substrate binding site). 3,10,11 According to kinetic studies on HMGRh, statins compete for the substrate HMG-CoA without affecting binding to the cofactor NADPH. 20 Hence, the substitution of an amino acid residue in the conserved sequences could possibly alter the a nity of the enzyme for its substrate HMG-CoA as well as for simvastatin and alpha-asarone, to demonstrate this, a molecular docking study was carried out.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, it has been proposed as an antifungal target. 10,11 The recombinant HMGRCg enzyme is available and its catalytic activity is still under study. Therefore, the aim of the current contribution was to generate, characterize, and overexpress ve point mutants of HMGRCg in order to explore the role of certain amino acid residues in the functioning of the catalytic domain of HMGRCg.…”

Section: Introductionmentioning

confidence: 99%

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Point Mutations In Candida Glabrata 3-Hydroxy-3-Methyl Glutaryl Coenzyme A Reductase (HMGRCg) An Antifungal Target, Decrease Enzymatic Activity And Substrate/Inhibitor Affinity

Andrade-Pavón

Fernández-Muñoz

González-Ibarra

et al. 2021

Preprint

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An alternative target for antifungal drugs is 3-hydroxy-3-methyl glutaryl coenzyme A reductase (HMGR), a key enzyme in the ergosterol biosynthesis pathway. The aim of this study was to obtain, purify, characterize, and overexpress five point mutations in highly conserved regions of the catalytic domain of Candida glabrata HMGR (HMGRCg) to explore the function of key amino acid residues. Glutamic acid (Glu) was substituted by glutamine in the E680Q mutant (at the dimerization site), Glu by glutamine in E711Q (at the substrate binding site), aspartic acid by alanine in D805A and methionine by arginine in M807R (the latter two at the cofactor binding site). A double mutation, E680Q-M807R, was also made. The in vitro enzymatic activity decreased significantly in all recombinant (versus wild-type) HMGRCg, and the in silico binding energy for simvastatin, alpha-asarone and the substrate HMG-CoA was also lower for the mutants. The lowest enzymatic activity and binding energy was displayed by E711Q, suggesting that Glu711 (in the substrate binding site) is an important residue for enzymatic activity. The double mutant HMGRCg E680Q-M807R exhibited the second lowest enzymatic activity. The current findings provide insights into the role of residues in the catalytic site of HMGRCg.

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Section: Discussionmentioning

confidence: 99%