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The Active Site Sulfenic Acid Ligand in Nitrile Hydratases Can Function as a Nucleophile
[Image: see text] Nitrile hydratase (NHase) catalyzes the hydration of nitriles to their corresponding commercially valuable amides at ambient temperatures and physiological pH. Several reaction mechanisms have been proposed for NHase enzymes; however, the source of the nucleophile remains a mystery...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical
Society
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3968781/ https://www.ncbi.nlm.nih.gov/pubmed/24383915 http://dx.doi.org/10.1021/ja410462j |
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author | Martinez, Salette Wu, Rui Sanishvili, Ruslan Liu, Dali Holz, Richard |
author_facet | Martinez, Salette Wu, Rui Sanishvili, Ruslan Liu, Dali Holz, Richard |
author_sort | Martinez, Salette |
collection | PubMed |
description | [Image: see text] Nitrile hydratase (NHase) catalyzes the hydration of nitriles to their corresponding commercially valuable amides at ambient temperatures and physiological pH. Several reaction mechanisms have been proposed for NHase enzymes; however, the source of the nucleophile remains a mystery. Boronic acids have been shown to be potent inhibitors of numerous hydrolytic enzymes due to the open shell of boron, which allows it to expand from a trigonal planar (sp(2)) form to a tetrahedral form (sp(3)). Therefore, we examined the inhibition of the Co-type NHase from Pseudonocardia thermophila JCM 3095 (PtNHase) by boronic acids via kinetics and X-ray crystallography. Both 1-butaneboronic acid (BuBA) and phenylboronic acid (PBA) function as potent competitive inhibitors of PtNHase. X-ray crystal structures for BuBA and PBA complexed to PtNHase were solved and refined at 1.5, 1.6, and 1.2 Å resolution. The resulting PtNHase–boronic acid complexes represent a “snapshot” of reaction intermediates and implicate the cysteine-sulfenic acid ligand as the catalytic nucleophile, a heretofore unknown role for the αCys(113)–OH sulfenic acid ligand. Based on these data, a new mechanism of action for the hydration of nitriles by NHase is presented. |
format | Online Article Text |
id | pubmed-3968781 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | American Chemical
Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-39687812015-01-02 The Active Site Sulfenic Acid Ligand in Nitrile Hydratases Can Function as a Nucleophile Martinez, Salette Wu, Rui Sanishvili, Ruslan Liu, Dali Holz, Richard J Am Chem Soc [Image: see text] Nitrile hydratase (NHase) catalyzes the hydration of nitriles to their corresponding commercially valuable amides at ambient temperatures and physiological pH. Several reaction mechanisms have been proposed for NHase enzymes; however, the source of the nucleophile remains a mystery. Boronic acids have been shown to be potent inhibitors of numerous hydrolytic enzymes due to the open shell of boron, which allows it to expand from a trigonal planar (sp(2)) form to a tetrahedral form (sp(3)). Therefore, we examined the inhibition of the Co-type NHase from Pseudonocardia thermophila JCM 3095 (PtNHase) by boronic acids via kinetics and X-ray crystallography. Both 1-butaneboronic acid (BuBA) and phenylboronic acid (PBA) function as potent competitive inhibitors of PtNHase. X-ray crystal structures for BuBA and PBA complexed to PtNHase were solved and refined at 1.5, 1.6, and 1.2 Å resolution. The resulting PtNHase–boronic acid complexes represent a “snapshot” of reaction intermediates and implicate the cysteine-sulfenic acid ligand as the catalytic nucleophile, a heretofore unknown role for the αCys(113)–OH sulfenic acid ligand. Based on these data, a new mechanism of action for the hydration of nitriles by NHase is presented. American Chemical Society 2014-01-02 2014-01-29 /pmc/articles/PMC3968781/ /pubmed/24383915 http://dx.doi.org/10.1021/ja410462j Text en Copyright © 2014 American Chemical Society |
spellingShingle | Martinez, Salette Wu, Rui Sanishvili, Ruslan Liu, Dali Holz, Richard The Active Site Sulfenic Acid Ligand in Nitrile Hydratases Can Function as a Nucleophile |
title | The Active
Site Sulfenic Acid Ligand in Nitrile Hydratases
Can Function as a Nucleophile |
title_full | The Active
Site Sulfenic Acid Ligand in Nitrile Hydratases
Can Function as a Nucleophile |
title_fullStr | The Active
Site Sulfenic Acid Ligand in Nitrile Hydratases
Can Function as a Nucleophile |
title_full_unstemmed | The Active
Site Sulfenic Acid Ligand in Nitrile Hydratases
Can Function as a Nucleophile |
title_short | The Active
Site Sulfenic Acid Ligand in Nitrile Hydratases
Can Function as a Nucleophile |
title_sort | active
site sulfenic acid ligand in nitrile hydratases
can function as a nucleophile |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3968781/ https://www.ncbi.nlm.nih.gov/pubmed/24383915 http://dx.doi.org/10.1021/ja410462j |
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