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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...

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Autores principales: 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
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
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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.
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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
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