Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675

Patrick E. Konold, Eunjin Yoon, Junghwa Lee, Samantha L. Allen, Prem P. Chapagain, Bernard S. Gerstman, Chola K. Regmi, Kiryl D. Piatkevich, Vladislav Verkhusha, Taiha Joo, Ralph Jimenez

Research output: Contribution to journalArticle

2 Citations (Scopus)

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/M41Q variant 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 S1 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.

Original languageEnglish (US)
Pages (from-to)3046-3051
Number of pages6
JournalJournal of Physical Chemistry Letters
Volume7
Issue number15
DOIs
StatePublished - Aug 4 2016

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Chromophores
chromophores
Hydrogen bonds
Fluorescence
proteins
Proteins
fluorescence
hydrogen
hydrogen bonds
Photoexcitation
Fluorescence spectroscopy
Ground state
Conformations
Molecular dynamics
Tuning
Derivatives
Imaging techniques
photoexcitation
blue shift
red shift

ASJC Scopus subject areas

  • Materials Science(all)

Cite this

Konold, P. E., Yoon, E., Lee, J., Allen, S. L., Chapagain, P. P., Gerstman, B. S., ... Jimenez, R. (2016). Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675. Journal of Physical Chemistry Letters, 7(15), 3046-3051. https://doi.org/10.1021/acs.jpclett.6b01172

Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675. / Konold, Patrick E.; Yoon, Eunjin; Lee, Junghwa; Allen, Samantha L.; Chapagain, Prem P.; Gerstman, Bernard S.; Regmi, Chola K.; Piatkevich, Kiryl D.; Verkhusha, Vladislav; Joo, Taiha; Jimenez, Ralph.

In: Journal of Physical Chemistry Letters, Vol. 7, No. 15, 04.08.2016, p. 3046-3051.

Research output: Contribution to journalArticle

Konold, PE, Yoon, E, Lee, J, Allen, SL, Chapagain, PP, Gerstman, BS, Regmi, CK, Piatkevich, KD, Verkhusha, V, Joo, T & Jimenez, R 2016, 'Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675', Journal of Physical Chemistry Letters, vol. 7, no. 15, pp. 3046-3051. https://doi.org/10.1021/acs.jpclett.6b01172
Konold, Patrick E. ; Yoon, Eunjin ; Lee, Junghwa ; Allen, Samantha L. ; Chapagain, Prem P. ; Gerstman, Bernard S. ; Regmi, Chola K. ; Piatkevich, Kiryl D. ; Verkhusha, Vladislav ; Joo, Taiha ; Jimenez, Ralph. / Fluorescence from Multiple Chromophore Hydrogen-Bonding States in the Far-Red Protein TagRFP675. In: Journal of Physical Chemistry Letters. 2016 ; Vol. 7, No. 15. pp. 3046-3051.
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