期刊
JOURNAL OF PHYSICAL CHEMISTRY B
卷 124, 期 33, 页码 7152-7165出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpcb.0c04943
关键词
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资金
- National Science Foundation (NSF) [MCB-1817837, MCB-1750637]
- EPSRC [EP/N033647/1]
- National Institutes of Health Chemistry-Biology Interface Training Grant [T32GM092714]
- Research Corporation for Science Advancement
- [EFOP-3.6.2-16-2017-00005]
- EPSRC [EP/N033647/1] Funding Source: UKRI
Flavoproteins are important blue light sensors in photobiology and play a key role in optogenetics. The characterization of their excited state structure and dynamics is thus an important objective. Here, we present a detailed study of excited state vibrational spectra of flavin mononucleotide (FMN), in solution and bound to the LOV-2 (Light-Oxygen-Voltage) domain of Avena saliva phototropin. Vibrational frequencies are determined for the optically excited singlet state and the reactive triplet state, through resonant ultrafast femtosecond stimulated Raman spectroscopy (FSRS). To assign the observed spectra, vibrational frequencies of the excited states are calculated using density functional theory, and both measurement and theory are applied to four different isotopologues of FMN. Excited state mode assignments are refined in both states, and their sensitivity to deuteration and protein environment are investigated. We show that resonant FSRS provides a useful tool for characterizing photoactive flavoproteins and is able to highlight chromophore localized modes and to record hydrogen/deuterium exchange.
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