Characterisation of NADH fluorescence properties under one-photon excitation with respect to temperature, pH and binding to lactate dehydrogenase
File(s)
Published version
Author(s)
Type
Journal Article
Abstract
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.
Date Issued
2021-05-15
Date Acceptance
2021-04-25
Citation
Optical Society of America Continuum, 2021, 4 (5), pp.1610-1625
Publisher
OSA Publishing
Start Page
1610
End Page
1625
Journal / Book Title
Optical Society of America Continuum
Volume
4
Issue
5
Copyright Statement
© 2021. 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.
distribution of this work must maintain attribution to the author(s) and the published article’s title, journal
citation, and DOI.
License URL
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
https://www.osapublishing.org/osac/fulltext.cfm?uri=osac-4-5-1610&id=450919
Grant Number
EP/I02770X/1
BB/E000495/1
Subjects
Science & Technology
Physical Sciences
Optics
NICOTINAMIDE ADENINE-DINUCLEOTIDE
MULTIPHOTON EXCITATION
REDUCED NICOTINAMIDES
INTRACELLULAR NADH
DECAY KINETICS
LIFETIME
DYNAMICS
NAD(P)H
CELLS
STATE
Publication Status
Published
Date Publish Online
2021-05-15