A fraction of NADH in solution is “dark”: Implications for metabolic sensing via fluorescence lifetime

Cao, Simin; Zhou, Zhongneng; Li, Haoyang; Jia, Menghui; Liu, Yangyi; Wang, Mengyu; Zhang, Mengjie; Zhang, Sanjun; Chen, Jacqueline; Xu, Jianhua; Knutson, Jay R.

doi:10.1016/j.cplett.2019.04.014

Cited by 14 publications

(35 citation statements)

References 39 publications

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“…Decay-associated spectra (DAS) and time-resolved evolution spectra (TRES) have been widely used for the study of ground-state heterogeneity, photochemical reaction, and solvation dynamics [ 13 , 14 , 15 , 16 ]. These spectra (with 10–15 nm resolution) are traditionally reconstructed by recording 10–20 kinetic traces I(t) at different wavelengths, which cover the whole emission region.…”

Section: Experimental Set-up and Considerationsmentioning

confidence: 99%

“…The ultrafast dynamics of reduced nicotinamide adenine dinucleotide (NADH) has been rarely studied. In our recent work, NADH spectral evolution was also found to show two distinct solvation dynamics terms on ps time scales [ 16 ]. Figure 7 gives the upconversion fluorescence decay transients at different emission wavelengths [ 17 ].…”

Section: Solvation Dynamics Of Biomolecules: Bulk Water and “Biolomentioning

confidence: 99%

“…Becker et al [ 27 ] also saw an ultrafast component with a lifetime of ~50 ps in their newly developed ultrafast HPM detectors. More recently, time-resolved fluorescence of free NADH in solution has been investigated using our ultraviolet upconversion spectrophotofluorometer (IRF < 350 fs) [ 16 ]. Figure 10 shows the novel decay associated spectra of free NADH in solution [ 16 ].…”

Section: Qssq In Biomoleculesmentioning

confidence: 99%

“…More recently, time-resolved fluorescence of free NADH in solution has been investigated using our ultraviolet upconversion spectrophotofluorometer (IRF < 350 fs) [ 16 ]. Figure 10 shows the novel decay associated spectra of free NADH in solution [ 16 ]. The DAS for the two longer lifetimes (244 ps and 690 ps) has positive amplitude at all measured wavelengths, representing two different self-quenched conformation of the folded molecules (ground-state heterogeneity).…”

Section: Qssq In Biomoleculesmentioning

confidence: 99%

“…From conventional photodetector studies, it was known that NADH free in solution exhibited short (~0.4 ns) decay times, but order of magnitude (2–8 ns) greater lifetimes were seen when it bound to cognate sites on target proteins. Our recent work showed the free molecule also exhibited a “dark” population, which, depending on detector, could hide 20–40% of the molecules from view [ 16 , 17 ]. Enzymatic binding abolished the quench.…”

Section: Introductionmentioning

confidence: 99%

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Ultrafast Fluorescence Spectroscopy via Upconversion and Its Applications in Biophysics

Cao

Zhao

et al. 2021

Molecules

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In this review, the experimental set-up and functional characteristics of single-wavelength and broad-band femtosecond upconversion spectrophotofluorometers developed in our laboratory are described. We discuss applications of this technique to biophysical problems, such as ultrafast fluorescence quenching and solvation dynamics of tryptophan, peptides, proteins, reduced nicotinamide adenine dinucleotide (NADH), and nucleic acids. In the tryptophan dynamics field, especially for proteins, two types of solvation dynamics on different time scales have been well explored: ~1 ps for bulk water, and tens of picoseconds for “biological water”, a term that combines effects of water and macromolecule dynamics. In addition, some proteins also show quasi-static self-quenching (QSSQ) phenomena. Interestingly, in our more recent work, we also find that similar mixtures of quenching and solvation dynamics occur for the metabolic cofactor NADH. In this review, we add a brief overview of the emerging development of fluorescent RNA aptamers and their potential application to live cell imaging, while noting how ultrafast measurement may speed their optimization.

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