Fluorescence dynamics of N-terminal Trp–Trp residues in polypeptide: intrinsic indicators for monitoring pH

Li, Lei; Hua, Yi; Chang, Mengfang; Cao, Xiaodan; Zhou, Zhongneng; Qin, Cuifang; Zhang, Sanjun; Pan, Haifeng; Chen, Yan; Xu, Jianhua

doi:10.1007/s11434-015-0942-5

Cited by 8 publications

(3 citation statements)

References 32 publications

(31 reference statements)

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“…The instrument response function was measured to be ∼440 ps from the Rayleigh scattering of SiO 2 nanoparticles in water. The details of this experimental setup have been described previously. − …”

Section: Methodsmentioning

confidence: 99%

Femtosecond Fluorescence Spectra of NADH in Solution: Ultrafast Solvation Dynamics

Cao

Liu

et al. 2020

J. Phys. Chem. B

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The ultrafast solvation dynamics of reduced nicotinamide adenine dinucleotide (NADH) free in solution has been investigated, using both a femtosecond upconversion spectrophotofluorometer and a picosecond time-correlated single-photon counting (TCSPC) apparatus. The familiar time constant of solvent relaxation originating in “bulk water” was found to be ∼1.4 ps, revealing ultrafast solvent reorientation upon excitation. We also found a slower spectral relaxation process with an apparent time of 27 ps, suggesting there could either be dissociable “biological water” hydration sites on the surface of NADH or internal dielectric rearrangements of the flexible solvated molecule on that timescale. In contrast, the femtosecond fluorescence anisotropy measurement revealed that rotational diffusion happened on two different timescales (3.6 ps (local) and 141 ps (tumbling)); thus, any dielectric rearrangement scenario for the 27 ps relaxation must occur without significant chromophore oscillator rotation. The coexistence of quasi-static self quenching (QSSQ) with the slower relaxation is also discussed.

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Section: Methodsmentioning

confidence: 99%

Femtosecond Fluorescence Spectra of NADH in Solution: Ultrafast Solvation Dynamics

Cao

Liu

et al. 2020

J. Phys. Chem. B

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“…More details about the system can be found elsewhere. 34 For constructing decay associated spectra (DAS), the fluorescence was scanned (in the range 420−560 nm for NADH−MDH and 420−500 nm for NADH−LDH) with 10 nm bandwidth and analyzed by a global fitting technique 35 MDH, ammonium sulfate suspension, was first centrifuged for 30 min (9000 rpm); then, the precipitate was redissolved in Tris−HCl buffer. After that, the solution was centrifugated for 30 min again to remove any insoluble particles.…”

Section: Methodsmentioning

confidence: 99%

“…The instrument response function was found to be ∼440 ps in width by measuring the Rayleigh scattering of excitation pulses in a suspension of 0.34 wt% SiO 2 nanoparticles in water. More details about the system can be found elsewhere . For constructing decay associated spectra (DAS), the fluorescence was scanned (in the range 420–560 nm for NADH–MDH and 420–500 nm for NADH–LDH) with 10 nm bandwidth and analyzed by a global fitting technique using a biexponential model.…”

Section: Experimental Sectionmentioning

confidence: 99%

Dehydrogenase Binding Sites Abolish the “Dark” Fraction of NADH: Implication for Metabolic Sensing via FLIM

Cao

Liu

et al. 2020

J. Phys. Chem. B

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The fluorescence of dinucleotide NADH has been exploited for decades to determine the redox state of cells and tissues in vivo and in vitro. Particularly, nanosecond (ns) fluorescence lifetime imaging microscopy (FLIM) of NADH (in free vs bound forms) has recently offered a label-free readout of mitochondrial function and allowed the different "pools" of NADH to be distinguished in living cells. In this study, the ultrafast fluorescence dynamics of NADH− dehydrogenase (MDH/LDH) complexes have been investigated by using both a femtosecond (fs) upconversion spectrophotofluorometer and a picosecond (ps) time-correlated single photon counting (TCSPC) apparatus. With these enhanced time-resolved tools, a few-picosecond decay process with a signatory spectrum was indeed found for bound NADH, and it can best be ascribed to the solvent relaxation originating in "bulk water". However, it is quite unlike our previously discovered ultrafast "dark" component (∼26 ps) that is prominent in free NADH (Chemical Physics Letters 2019, 726, 18−21). For these two critical protein-bound NADH exemplars, the decay transients lack the ultrafast quenching that creates the "dark" subpopulation of free NADH. Therefore, we infer that the apparent ratio of free to bound NADH recovered by ordinary (>50 ps) FLIM methods may be low, since the "dark" molecule subpopulation (lifetime too short for conventional FLIM), which effectively hides about a quarter of free molecules, is not present in the dehydrogenase-bound state.

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Fluorescence Dynamics of N-Terminal Tryptofan-X Residues in Polypeptide: pH Response

Cao

Zhou

et al. 2017

J Appl Spectrosc

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Fluorescence dynamics of N-terminal Trp–Trp residues in polypeptide: intrinsic indicators for monitoring pH

Cited by 8 publications

References 32 publications

Femtosecond Fluorescence Spectra of NADH in Solution: Ultrafast Solvation Dynamics

Femtosecond Fluorescence Spectra of NADH in Solution: Ultrafast Solvation Dynamics

Dehydrogenase Binding Sites Abolish the “Dark” Fraction of NADH: Implication for Metabolic Sensing via FLIM

Fluorescence Dynamics of N-Terminal Tryptofan-X Residues in Polypeptide: pH Response

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