Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675
SCIE
SCOPUS
- Title
- Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675
- Authors
- Patrick Konold; Eunjin Yoon; Lee, J; Allen, SL; Chapagain, PP; Gerstman, BS; Regmi, CK; Piatkevich, KD; Verkhusha, VV; Joo, T; Jimenez, R
- Date Issued
- 2016-08-04
- Publisher
- American Chemical Society
- Abstract
- Far-red fluorescent proteins are critical for in vivo imaging applications, but the relative importance of structure versus dynamics in generating large Stokes-shifted emission is unclear. The unusually red-shifted emission of TagRFP675, a derivative of mKate, has been attributed to the multiple hydrogen bonds with the chromophore N-acylimine carbonyl. We characterized TagRFP675 and point mutants designed to perturb these hydrogen bonds with spectrally resolved transient grating and time-resolved fluorescence (TRF) spectroscopies supported by molecular dynamics simulations. TRF results for TagRFP675 and the mKate/M41Qvariant show picosecond time scale red-shifts followed by nanosecond time blue-shifts. Global analysis of the TRF spectra reveals spectrally distinct emitting states that do not interconvert during the S-1 lifetime. These dynamics originate from photoexcitation of a mixed ground-state population of acylimine hydrogen bond conformers. Strategically tuning the chromophore environment in TagRFP675 might stabilize the most red-shifted conformation and result in a variant with a larger Stokes shift.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/37633
- DOI
- 10.1021/ACS.JPCLETT.6B01172
- ISSN
- 1948-7185
- Article Type
- Article
- Citation
- Journal of Physical Chemistry Letters, vol. 7, no. 15, page. 3046 - 3051, 2016-08-04
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