Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays

l‐Ascorbate (l‐Asc) is often added to assays with isolated Fe(II)‐ and 2‐oxoglutarate (2OG)‐dependent oxygenases to enhance activity. l‐Asc is proposed to be important in catalysis by some 2OG oxygenases in vivo. We report observations on the nonenzymatic conversion of 2OG to succinate, which is med...

Descripción completa

Detalles Bibliográficos
Autores principales: Khan, Amjad, Schofield, Christopher J., Claridge, Timothy D. W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Communications
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693218/
https://www.ncbi.nlm.nih.gov/pubmed/32478965
http://dx.doi.org/10.1002/cbic.202000185
_version_ 1783614693078728704
author Khan, Amjad
Schofield, Christopher J.
Claridge, Timothy D. W.
author_facet Khan, Amjad
Schofield, Christopher J.
Claridge, Timothy D. W.
author_sort Khan, Amjad
collection PubMed
description l‐Ascorbate (l‐Asc) is often added to assays with isolated Fe(II)‐ and 2‐oxoglutarate (2OG)‐dependent oxygenases to enhance activity. l‐Asc is proposed to be important in catalysis by some 2OG oxygenases in vivo. We report observations on the nonenzymatic conversion of 2OG to succinate, which is mediated by hydrogen peroxide generated by the reaction of l‐Asc and dioxygen. Slow nonenzymatic oxidation of 2OG to succinate occurs with some, but not all, other reducing agents commonly used in 2OG oxygenase assays. We intend these observations will help in the robust assignment of substrates and inhibitors for 2OG oxygenases.
format Online
Article
Text
id pubmed-7693218
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-76932182020-12-11 Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays Khan, Amjad Schofield, Christopher J. Claridge, Timothy D. W. Chembiochem Communications l‐Ascorbate (l‐Asc) is often added to assays with isolated Fe(II)‐ and 2‐oxoglutarate (2OG)‐dependent oxygenases to enhance activity. l‐Asc is proposed to be important in catalysis by some 2OG oxygenases in vivo. We report observations on the nonenzymatic conversion of 2OG to succinate, which is mediated by hydrogen peroxide generated by the reaction of l‐Asc and dioxygen. Slow nonenzymatic oxidation of 2OG to succinate occurs with some, but not all, other reducing agents commonly used in 2OG oxygenase assays. We intend these observations will help in the robust assignment of substrates and inhibitors for 2OG oxygenases. John Wiley and Sons Inc. 2020-08-18 2020-10-15 /pmc/articles/PMC7693218/ /pubmed/32478965 http://dx.doi.org/10.1002/cbic.202000185 Text en © 2020 The Authors. Published by Wiley-VCH GmbH This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Communications
Khan, Amjad
Schofield, Christopher J.
Claridge, Timothy D. W.
Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title_full Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title_fullStr Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title_full_unstemmed Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title_short Reducing Agent‐Mediated Nonenzymatic Conversion of 2‐Oxoglutarate to Succinate: Implications for Oxygenase Assays
title_sort reducing agent‐mediated nonenzymatic conversion of 2‐oxoglutarate to succinate: implications for oxygenase assays
topic Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693218/
https://www.ncbi.nlm.nih.gov/pubmed/32478965
http://dx.doi.org/10.1002/cbic.202000185
work_keys_str_mv AT khanamjad reducingagentmediatednonenzymaticconversionof2oxoglutaratetosuccinateimplicationsforoxygenaseassays
AT schofieldchristopherj reducingagentmediatednonenzymaticconversionof2oxoglutaratetosuccinateimplicationsforoxygenaseassays
AT claridgetimothydw reducingagentmediatednonenzymaticconversionof2oxoglutaratetosuccinateimplicationsforoxygenaseassays

Ejemplares similares