S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic

S-adenosyl-L-homocysteine (SAH) hydrolases (SAHases) are involved in the regulation of methylation reactions in many organisms and are thus crucial for numerous cellular functions. Consequently, their dysregulation is associated with severe health problems. The SAHase-catalyzed reaction is reversibl...

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Main Authors: Lyn L. Kailing, Daniela Bertinetti, Caroline E. Paul, Tomasz Manszewski, Mariusz Jaskolski, Friedrich W. Herberg, Ioannis V. Pavlidis
Format: Article
Language:English
Published: Frontiers Media S.A. 2018-03-01
Series:Frontiers in Microbiology
Subjects:
Online Access:http://journal.frontiersin.org/article/10.3389/fmicb.2018.00505/full
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spelling doaj-2fb79261d62b434fb4d1c54e2fb12c032020-11-24T22:25:50ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-03-01910.3389/fmicb.2018.00505345870S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor BiomimeticLyn L. Kailing0Daniela Bertinetti1Caroline E. Paul2Tomasz Manszewski3Mariusz Jaskolski4Mariusz Jaskolski5Friedrich W. Herberg6Ioannis V. Pavlidis7Ioannis V. Pavlidis8Department of Biochemistry, University of Kassel, Kassel, GermanyDepartment of Biochemistry, University of Kassel, Kassel, GermanyLaboratory of Organic Chemistry, Wageningen University & Research, Wageningen, NetherlandsCenter for Biocrystallographic Research, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, PolandCenter for Biocrystallographic Research, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, PolandDepartment of Crystallography, Faculty of Chemistry, Adam Mickiewicz University in Poznań, Poznań, PolandDepartment of Biochemistry, University of Kassel, Kassel, GermanyDepartment of Biochemistry, University of Kassel, Kassel, GermanyDepartment of Chemistry, University of Crete, Heraklion, GreeceS-adenosyl-L-homocysteine (SAH) hydrolases (SAHases) are involved in the regulation of methylation reactions in many organisms and are thus crucial for numerous cellular functions. Consequently, their dysregulation is associated with severe health problems. The SAHase-catalyzed reaction is reversible and both directions depend on the redox activity of nicotinamide adenine dinucleotide (NAD+) as a cofactor. Therefore, nicotinamide cofactor biomimetics (NCB) are a promising tool to modulate SAHase activity. In the present in vitro study, we investigated 10 synthetic truncated NAD+ analogs against a SAHase from the root-nodulating bacterium Bradyrhizobium elkanii. Among this set of analogs, one was identified to inhibit the SAHase in both directions. Isothermal titration calorimetry (ITC) and crystallography experiments suggest that the inhibitory effect is not mediated by a direct interaction with the protein. Neither the apo-enzyme (i.e., deprived of the natural cofactor), nor the holo-enzyme (i.e., in the NAD+-bound state) were found to bind the inhibitor. Yet, enzyme kinetics point to a non-competitive inhibition mechanism, where the inhibitor acts on both, the enzyme and enzyme-SAH complex. Based on our experimental results, we hypothesize that the NCB inhibits the enzyme via oxidation of the enzyme-bound NADH, which may be accessible through an open molecular gate, leaving the enzyme stalled in a configuration with oxidized cofactor, where the reaction intermediate can be neither converted nor released. Since the reaction mechanism of SAHase is quite uncommon, this kind of inhibition could be a viable pharmacological route, with a low risk of off-target effects. The NCB presented in this work could be used as a template for the development of more potent SAHase inhibitors.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00505/fullS-adenosyl-L-homocysteine hydrolaseinhibitionnicotinamide cofactorbiomimeticcrystallography
collection DOAJ
language English
format Article
sources DOAJ
author Lyn L. Kailing
Daniela Bertinetti
Caroline E. Paul
Tomasz Manszewski
Mariusz Jaskolski
Mariusz Jaskolski
Friedrich W. Herberg
Ioannis V. Pavlidis
Ioannis V. Pavlidis
spellingShingle Lyn L. Kailing
Daniela Bertinetti
Caroline E. Paul
Tomasz Manszewski
Mariusz Jaskolski
Mariusz Jaskolski
Friedrich W. Herberg
Ioannis V. Pavlidis
Ioannis V. Pavlidis
S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
Frontiers in Microbiology
S-adenosyl-L-homocysteine hydrolase
inhibition
nicotinamide cofactor
biomimetic
crystallography
author_facet Lyn L. Kailing
Daniela Bertinetti
Caroline E. Paul
Tomasz Manszewski
Mariusz Jaskolski
Mariusz Jaskolski
Friedrich W. Herberg
Ioannis V. Pavlidis
Ioannis V. Pavlidis
author_sort Lyn L. Kailing
title S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
title_short S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
title_full S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
title_fullStr S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
title_full_unstemmed S-Adenosyl-L-Homocysteine Hydrolase Inhibition by a Synthetic Nicotinamide Cofactor Biomimetic
title_sort s-adenosyl-l-homocysteine hydrolase inhibition by a synthetic nicotinamide cofactor biomimetic
publisher Frontiers Media S.A.
series Frontiers in Microbiology
issn 1664-302X
publishDate 2018-03-01
description S-adenosyl-L-homocysteine (SAH) hydrolases (SAHases) are involved in the regulation of methylation reactions in many organisms and are thus crucial for numerous cellular functions. Consequently, their dysregulation is associated with severe health problems. The SAHase-catalyzed reaction is reversible and both directions depend on the redox activity of nicotinamide adenine dinucleotide (NAD+) as a cofactor. Therefore, nicotinamide cofactor biomimetics (NCB) are a promising tool to modulate SAHase activity. In the present in vitro study, we investigated 10 synthetic truncated NAD+ analogs against a SAHase from the root-nodulating bacterium Bradyrhizobium elkanii. Among this set of analogs, one was identified to inhibit the SAHase in both directions. Isothermal titration calorimetry (ITC) and crystallography experiments suggest that the inhibitory effect is not mediated by a direct interaction with the protein. Neither the apo-enzyme (i.e., deprived of the natural cofactor), nor the holo-enzyme (i.e., in the NAD+-bound state) were found to bind the inhibitor. Yet, enzyme kinetics point to a non-competitive inhibition mechanism, where the inhibitor acts on both, the enzyme and enzyme-SAH complex. Based on our experimental results, we hypothesize that the NCB inhibits the enzyme via oxidation of the enzyme-bound NADH, which may be accessible through an open molecular gate, leaving the enzyme stalled in a configuration with oxidized cofactor, where the reaction intermediate can be neither converted nor released. Since the reaction mechanism of SAHase is quite uncommon, this kind of inhibition could be a viable pharmacological route, with a low risk of off-target effects. The NCB presented in this work could be used as a template for the development of more potent SAHase inhibitors.
topic S-adenosyl-L-homocysteine hydrolase
inhibition
nicotinamide cofactor
biomimetic
crystallography
url http://journal.frontiersin.org/article/10.3389/fmicb.2018.00505/full
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