Role of Local Electric Field in Controlling Fluorescence Quantum Yield of Red Fluorescent Proteins

2019 ◽  
Author(s):  
Mikhail Drobizhev ◽  
J. Nathan Scott ◽  
Patrik R. Callis ◽  
Rosana S. Molina ◽  
Thomas E. Hughes
Keyword(s):  
Quantum Yield ◽  
Electric Field ◽  
Fluorescence Quantum Yield ◽  
Fluorescent Proteins ◽  
Local Electric Field ◽  
Red Fluorescent Proteins

Ultrafast spectroscopy of biliverdin dimethyl ester in solution: pathways of excited-state depopulation

2020 ◽  
Vol 22 (35) ◽  
pp. 19903-19912
Author(s):  
Yangyi Liu ◽  
Zhuang Chen ◽  
Xueli Wang ◽  
Simin Cao ◽  
Jianhua Xu ◽  
...  
Keyword(s):  
Excited State ◽  
Quantum Yield ◽  
Quantum Efficiency ◽  
Fluorescence Quantum Yield ◽  
Ultrafast Spectroscopy ◽  
Fluorescent Proteins ◽  
Dimethyl Ester ◽  
Bile Pigments ◽  
Enhanced Fluorescence ◽  
Red Fluorescent Proteins

Biliverdin and its dimethyl ester derivatives are bile pigments with very low fluorescence quantum yield in solution, but naturally serve as chromophores in far-red fluorescent proteins with three orders of magnitude enhanced fluorescence quantum efficiency.

scholarly journals Local Electric Field Controls Fluorescence Quantum Yield of Red and Far-Red Fluorescent Proteins

2021 ◽  
Vol 8 ◽  
Author(s):  
Mikhail Drobizhev ◽  
Rosana S. Molina ◽  
Patrik R. Callis ◽  
J. Nathan Scott ◽  
Gerard G. Lambert ◽  
...  
Keyword(s):  
Charge Transfer ◽  
Electric Field ◽  
Fluorescence Quantum Yield ◽  
Energy Gap ◽  
Intramolecular Charge Transfer ◽  
Fluorescent Proteins ◽  
Quantum Yields ◽  
Nonradiative Relaxation ◽  
Red Fluorescent Proteins ◽  
Energy Gap Law

Genetically encoded probes with red-shifted absorption and fluorescence are highly desirable for imaging applications because they can report from deeper tissue layers with lower background and because they provide additional colors for multicolor imaging. Unfortunately, red and especially far-red fluorescent proteins have very low quantum yields, which undermines their other advantages. Elucidating the mechanism of nonradiative relaxation in red fluorescent proteins (RFPs) could help developing ones with higher quantum yields. Here we consider two possible mechanisms of fast nonradiative relaxation of electronic excitation in RFPs. The first, known as the energy gap law, predicts a steep exponential drop of fluorescence quantum yield with a systematic red shift of fluorescence frequency. In this case the relaxation of excitation occurs in the chromophore without any significant changes of its geometry. The second mechanism is related to a twisted intramolecular charge transfer in the excited state, followed by an ultrafast internal conversion. The chromophore twisting can strongly depend on the local electric field because the field can affect the activation energy. We present a spectroscopic method of evaluating local electric fields experienced by the chromophore in the protein environment. The method is based on linear and two-photon absorption spectroscopy, as well as on quantum-mechanically calculated parameters of the isolated chromophore. Using this method, which is substantiated by our molecular dynamics simulations, we obtain the components of electric field in the chromophore plane for seven different RFPs with the same chromophore structure. We find that in five of these RFPs, the nonradiative relaxation rate increases with the strength of the field along the chromophore axis directed from the center of imidazolinone ring to the center of phenolate ring. Furthermore, this rate depends on the corresponding electrostatic energy change (calculated from the known fields and charge displacements), in quantitative agreement with the Marcus theory of charge transfer. This result supports the dominant role of the twisted intramolecular charge transfer mechanism over the energy gap law for most of the studied RFPs. It provides important guidelines of how to shift the absorption wavelength of an RFP to the red, while keeping its brightness reasonably high.

scholarly journals Quantitative Determination of Dark Chromophore Population Explains the Apparent Low Quantum Yield of Red Fluorescent Proteins

2020 ◽  
Vol 124 (8) ◽  
pp. 1383-1391 ◽  
Author(s):  
Jord C. Prangsma ◽  
Robert Molenaar ◽  
Laura van Weeren ◽  
Daphne S. Bindels ◽  
Lindsay Haarbosch ◽  
...  
Keyword(s):  
Quantum Yield ◽  
Quantitative Determination ◽  
Fluorescent Proteins ◽  
Red Fluorescent Proteins

scholarly journals Generation of bright monomeric red fluorescent proteins via computational design of enhanced chromophore packing

2022 ◽  
Author(s):  
Sandrine Legault ◽  
Derek Paco Fraser-Halberg ◽  
Ralph McAnelly ◽  
Matthew G Eason ◽  
Michael Thompson ◽  
...  
Keyword(s):  
Quantum Yield ◽  
Protein Design ◽  
Fluorescent Proteins ◽  
Computational Design ◽  
Computational Protein Design ◽  
Biological Research ◽  
Widespread Application ◽  
Red Fluorescent Proteins

Red fluorescent proteins (RFPs) have found widespread application in chemical and biological research due to their longer emission wavelengths. Here, we use computational protein design to increase the quantum yield...

scholarly journals Quantification of reactive oxygen species production by the red fluorescent proteins KillerRed, SuperNova and mCherry

2019 ◽  
Author(s):  
John O. Onukwufor ◽  
Adam J. Trewin ◽  
Timothy M. Baran ◽  
Anmol Almast ◽  
Thomas H. Foster ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Quantum Yield ◽  
Singlet Oxygen ◽  
Fluorescent Proteins ◽  
Reactive Oxygen ◽  
Quantum Yields ◽  
Type I ◽  
Oxygen Species ◽  
Redox Biology ◽  
Red Fluorescent Proteins

ABSTRACTFluorescent proteins can generate reactive oxygen species (ROS) upon absorption of photons via type I and II photosensitization mechanisms. The red fluorescent proteins KillerRed and SuperNova are phototoxic proteins engineered to generate ROS and are used in a variety of biological applications. However, their relative quantum yields and rates of ROS production are unclear, which has limited the interpretation of their effects when used in biological systems. We cloned and purified KillerRed, SuperNova, and mCherry - a related red fluorescent protein not typically considered a photosensitizer - and measured the superoxide (O2•-) and singlet oxygen (1O2) quantum yields with irradiation at 561 nm. The formation of the O2•--specific product 2-hydroxyethidium (2-OHE+) was quantified via HPLC separation with fluorescence detection. Relative to a reference photosensitizer, Rose Bengal, the O2•- quantum yield (ΦO2•-) of SuperNova was determined to be 0.00150, KillerRed was 0.00097, and mCherry 0.00120. At an excitation fluence of 916.5 J/cm2 and matched absorption at 561 nm, SuperNova, KillerRed and mCherry made 3.81, 2.38 and 1.65 μM O2•-/min, respectively. Using the probe Singlet Oxygen Sensor Green (SOSG), we ascertained the 1O2 quantum yield (Φ1O2) for SuperNova to be 0.0220, KillerRed 0.0076, and mCherry 0.0057. These photosensitization characteristics of SuperNova, KillerRed and mCherry improve our understanding of fluorescent proteins and are pertinent for refining their use as tools to advance our knowledge of redox biology.GRAPHICAL ABSTRACT

POSSIBLE APPLICATION OF METAL SENSITIVE RED FLUORESCENT PROTEINS IN ENVIRONMENTAL MONITORING

2012 ◽  
Vol 11 (1) ◽  
pp. 193-198 ◽  
Author(s):  
Laszlo Szilagyi ◽  
Maria Szabo (Palfi) ◽  
Judit Petres ◽  
Ildiko Miklossy ◽  
Beata Abraham ◽  
...  
Keyword(s):  
Environmental Monitoring ◽  
Fluorescent Proteins ◽  
Red Fluorescent Proteins
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