Cargando…
Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase
Reduced nicotinamide adenine dinucleotide (NADH) is the principal electron donor in glycolysis and oxidative metabolism and is thus recognized as a key biomarker for probing metabolic state. While the fluorescence characteristics of NADH have been investigated extensively, there are discrepancies in...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Optical Society of America
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8367293/ https://www.ncbi.nlm.nih.gov/pubmed/34458690 http://dx.doi.org/10.1364/OSAC.423082 |
_version_ | 1783739039015239680 |
---|---|
author | Cannon, Taylor M. Lagarto, Joao L. Dyer, Benjamin T. Garcia, Edwin Kelly, Douglas J. Peters, Nicholas S. Lyon, Alexander R. French, Paul M. W. Dunsby, Chris |
author_facet | Cannon, Taylor M. Lagarto, Joao L. Dyer, Benjamin T. Garcia, Edwin Kelly, Douglas J. Peters, Nicholas S. Lyon, Alexander R. French, Paul M. W. Dunsby, Chris |
author_sort | Cannon, Taylor M. |
collection | PubMed |
description | Reduced nicotinamide adenine dinucleotide (NADH) is the principal electron donor in glycolysis and oxidative metabolism and is thus recognized as a key biomarker for probing metabolic state. While the fluorescence characteristics of NADH have been investigated extensively, there are discrepancies in the published data due to diverse experimental conditions, instrumentation and microenvironmental parameters that can affect NADH fluorescence. Using a cuvette-based time-resolved spectrofluorimeter employing one-photon excitation at 375 nm, we characterized the fluorescence intensity, lifetime, spectral response, anisotropy and time-resolved anisotropy of NADH in aqueous solution under varying microenvironmental conditions, namely temperature, pH, and binding to lactate dehydrogenase (LDH). Our results demonstrate how temperature, pH, and binding partners each impact the fluorescence signature of NADH and highlight the complexity of the fluorescence data when different parameters produce competing effects. We hope that the data presented in this study will provide a reference for potential sources of variation in experiments measuring NADH fluorescence. |
format | Online Article Text |
id | pubmed-8367293 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Optical Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-83672932021-08-27 Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase Cannon, Taylor M. Lagarto, Joao L. Dyer, Benjamin T. Garcia, Edwin Kelly, Douglas J. Peters, Nicholas S. Lyon, Alexander R. French, Paul M. W. Dunsby, Chris OSA Contin Article Reduced nicotinamide adenine dinucleotide (NADH) is the principal electron donor in glycolysis and oxidative metabolism and is thus recognized as a key biomarker for probing metabolic state. While the fluorescence characteristics of NADH have been investigated extensively, there are discrepancies in the published data due to diverse experimental conditions, instrumentation and microenvironmental parameters that can affect NADH fluorescence. Using a cuvette-based time-resolved spectrofluorimeter employing one-photon excitation at 375 nm, we characterized the fluorescence intensity, lifetime, spectral response, anisotropy and time-resolved anisotropy of NADH in aqueous solution under varying microenvironmental conditions, namely temperature, pH, and binding to lactate dehydrogenase (LDH). Our results demonstrate how temperature, pH, and binding partners each impact the fluorescence signature of NADH and highlight the complexity of the fluorescence data when different parameters produce competing effects. We hope that the data presented in this study will provide a reference for potential sources of variation in experiments measuring NADH fluorescence. Optical Society of America 2021-05-10 /pmc/articles/PMC8367293/ /pubmed/34458690 http://dx.doi.org/10.1364/OSAC.423082 Text en Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI. https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Cannon, Taylor M. Lagarto, Joao L. Dyer, Benjamin T. Garcia, Edwin Kelly, Douglas J. Peters, Nicholas S. Lyon, Alexander R. French, Paul M. W. Dunsby, Chris Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title | Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title_full | Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title_fullStr | Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title_full_unstemmed | Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title_short | Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase |
title_sort | characterization of nadh fluorescence properties under one-photon excitation with respect to temperature, ph, and binding to lactate dehydrogenase |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8367293/ https://www.ncbi.nlm.nih.gov/pubmed/34458690 http://dx.doi.org/10.1364/OSAC.423082 |
work_keys_str_mv | AT cannontaylorm characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT lagartojoaol characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT dyerbenjamint characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT garciaedwin characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT kellydouglasj characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT petersnicholass characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT lyonalexanderr characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT frenchpaulmw characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase AT dunsbychris characterizationofnadhfluorescencepropertiesunderonephotonexcitationwithrespecttotemperaturephandbindingtolactatedehydrogenase |