Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase.
Characterization of NADH fluorescence properties under one-photon excitation with respect to temperature, pH, and binding to lactate dehydrogenase.
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DOI：
10.1364/osac.423082
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发表时间：
2021-05-15
期刊：
影响因子：
1.6
通讯作者：
Dunsby C
中科院分区：
文献类型：
--
作者：
Cannon TM;Lagarto JL;Dyer BT;Garcia E;Kelly DJ;Peters NS;Lyon AR;French PMW;Dunsby C
还原烟酰胺腺嘌呤二核苷酸（NADH）是糖酵解和氧化代谢的主要电子供体，因此被认为是探测代谢状态的关键生物标志物。虽然对NADH的荧光特性进行了广泛的研究，但由于不同的实验条件、仪器和微环境参数会影响NADH的荧光，因此发表的数据存在差异。利用基于比色管的时间分辨荧光仪，在375 nm处采用单光子激发，我们表征了不同微环境条件下，即温度、pH和与乳酸脱氢酶（LDH）的结合，水溶液中NADH的荧光强度、寿命、光谱响应、各向异性和时间分辨各向异性。我们的研究结果展示了温度、pH和结合伙伴如何影响NADH的荧光特征，并突出了当不同参数产生竞争效应时荧光数据的复杂性。我们希望本研究提供的数据能够为NADH荧光测量实验中潜在的变异源提供参考。
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.