Origin, Nature, and Fate of the Fluorescent State of the Green Fluorescent Protein Chromophore at the CASPT2//CASSCF Resolution

Martı́n, M. Elena; Negri, Fabrizia; Olivucci, Massimo

doi:10.1021/ja037278m

Cited by 314 publications

(503 citation statements)

References 30 publications

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“…72). It is notable that this differs qualitatively from the previous CAS(12e,11o) result with twostate averaging, 68 which suggests that the choice of the states to be averaged is as important as the choice of the active space.…”

Section: Conical Intersections Of Phbdicontrasting

confidence: 90%

“…Anionic pHBDI is considered to be an emitting species of the green fluorescent protein (GFP) and its variants. 6,[66][67][68][69][70][71][72][73][74][75][76] While GFP exhibits strong fluorescence with a lifetime on the order of nanoseconds, 6,66,[68][69][70][71][72][73] the nonadiabatic transition is known to occur in about a few picoseconds when the chromophore is not embedded in the protein environment. 77 As a resonant monomethine dye, it is widely accepted that the anionic GFP chromophore undergoes nonadiabatic transitions when the chromophore is twisted along the bridge.…”

Section: Conical Intersections Of Phbdimentioning

confidence: 99%

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Analytical Derivative Coupling for Multistate CASPT2 Theory

Park

Shiozaki

2017

J. Chem. Theory Comput.

139

169

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The probability of non-radiative transitions in photochemical dynamics is determined by the derivative couplings, the couplings between different electronic states through the nuclear degrees of freedom. Efficient and accurate evaluation of the derivative couplings is, therefore, of central importance to realize reliable computer simulations of photochemical reactions. In this work, the derivative couplings for multistate multireference second-order perturbation theory (MS-CASPT2) and its 'extended' variant (XMS-CASPT2) are studied, in which we present an algorithm for their analytical evaluation. The computational costs for evaluating the derivative couplings are essentially the same as those for calculating the nuclear energy gradients. The geometries and energies calculated with XMS-CASPT2 for small molecules at minimum energy conical intersections (MECIs) are in good agreement with those computed by multireference configuration interaction. As numerical examples, MECIs are optimized using XMS-CASPT2 for stilbene and a GFP model chromophore (the 4-para-hydroxybenzylidene-1,2-dimethyl-imidazolin-5-one anion).

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Section: Conical Intersections Of Phbdicontrasting

confidence: 90%

Section: Conical Intersections Of Phbdimentioning

confidence: 99%

Analytical Derivative Coupling for Multistate CASPT2 Theory

Park

Shiozaki

2017

J. Chem. Theory Comput.

139

169

View full text Add to dashboard Cite

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“…To this end, we adopted high-level quantum mechanical computations on the chemically very similar chromophore of GFP (26). When starting the simulation from the first electronically excited energy level of the asFP595 off state, the bridging methine group spontaneously rotated toward H197 within 100-500 fs through a bottom hula twist (HT bot ) (see Fig.…”

Section: Resultsmentioning

confidence: 99%

Structure and mechanism of the reversible photoswitch of a fluorescent protein

Andresen

Wahl

Stiel

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

256

341

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Proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state bear enormous potential in diverse fields, such as data storage, in vivo protein tracking, and subdiffraction resolution light microscopy. However, these proteins could hitherto not live up to their full potential because the molecular switching mechanism is not resolved. Here, we clarify the molecular photoswitching mechanism of asFP595, a green fluorescent protein (GFP)-like protein that can be transferred from a nonfluorescent ''off'' to a fluorescent ''on'' state and back again, by green and blue light, respectively. To this end, we establish reversible photoswitching of fluorescence in whole protein crystals and show that the switching kinetics in the crystal is identical with that in solution. Subsequent x-ray analysis demonstrated that upon the absorption of a green photon, the chromophore isomerizes from a trans (off) to a cis (on) state. Molecular dynamics calculations suggest that isomerization occurs through a bottom hula twist mechanism with concomitant rotation of both bonds of the chromophoric methine ring bridge. This insight into the switching mechanism should facilitate the targeted design of photoswitchable proteins. Reversible photoswitching of the protein chromophore system within intact crystals also constitutes a step toward the use of fluorescent proteins in three-dimensional data recording.photoisomerization ͉ asCP ͉ photochromism ͉ optical bistability ͉ asulCP F luorescent proteins have been widely used as genetically encodable tags to monitor protein localizations and dynamics in live cells (1-3). Recently, novel GFP-like fluorescent proteins have been discovered (4-6) that can be reversibly photoswitched between a fluorescent (on) and nonfluorescent (off) state, that is, they are optically bistable and fluorescent. This feature is remarkable, because the reversible photoswitching occurring in photochromic organic compounds, such as in fulgides and diarylethenes, is usually not accompanied by fluorescence (7). Therefore, not surprisingly, these proteins hold great promise in many areas of science reaching out far beyond their prominent use as triggerable protein markers in live cells. For example, the reversible photoswitching of fluorescent markers should provide nanoscale resolution in fluorescence microscopy by using lenses and regular illumination, which was hardly conceivable only a few years ago (8-10). As fluorescence can be sensitively read out from a bulky crystal, the prospect of erasable three-dimensional data storage is equally intriguing.The GFP-like protein asFP595 (asCP or asulCP) from the sea anemone Anemonia sulcata is such a protein: It can be transferred by green light from a nonfluorescent off into a fluorescent on state from which it reverts back eventually, but this transition can also be promptly stimulated by gentle irradiation with blue light (6). The ''on-off'' cycle can be repeated many times. However, with its low quantum yield (Ͻ0.001, ref. 6) and comparatively slo...

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“…[6][7][8] Although GFP was discovered more than 20 years ago, the photophysics of p-HBI and of its mutants is still intensively investigated mainly because of the spectacular decrease of its fluorescence quantum yield, Φ fl , and lifetime, τ, taking place when going from the protein (Φ fl ) 0.79, 9 τ ) 3.3 ns) 8,10 to liquid solutions (Φ fl < 10 -3 , 11 τ < 1 ps). [12][13][14] In this respect, the photophysical properties of the synthetic dimethyl derivative of the GFP chromophore (p-HBDI) 13,[15][16][17][18][19][20][21][22][23][24][25][26] and other related model systems 24,25,[27][28][29][30][31][32] have been extensively investigated.…”

Section: Introductionmentioning

confidence: 99%

Tuning the Excited-State Dynamics of GFP-Inspired Imidazolone Derivatives

et al. 2009

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The excited-state dynamics of five derivatives of the GFP-chromophore, which differ by the position and nature of their substituents, has been investigated in solvents of various viscosity and polarity and in rigid media using femtosecond-resolved spectroscopy. In polar solvents of low viscosity, like acetonitrile or methanol, the fluorescence decays of all compounds are multiexponential, with average lifetimes of the order of a few picoseconds, whereas in rigid matrices (polymer films and low temperature glasses), they are single exponential with lifetimes of the order of a few nanoseconds and fluorescence quantum yields close to unity. Global analysis of the fluorescence decays recorded at several wavelengths and of the transient absorption spectra reveals the presence of several excited-state populations with slightly different fluorescence and absorption spectra and with distinct lifetimes. These populations are attributed to the existence of multiple ground-state conformers. From the analysis of the dependence of the excited-state dynamics on the solvent and on the nature of the substituents, it follows that the nonradiative deactivation of all these excited chromophores involves an intramolecular coordinate with large amplitude motion. However, depending on the solvent and substituent, additional channels, namely, inter-and intramolecular hydrogen bond assisted nonradiative deactivation, are operative. This allows tuning of the excited-state lifetime of the chromophore. Finally, an ultrafast photoinduced intramolecular charge transfer is observed in polar solvents with one derivative bearing a dimethylaminophenyl substituent.

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Origin, Nature, and Fate of the Fluorescent State of the Green Fluorescent Protein Chromophore at the CASPT2//CASSCF Resolution

Cited by 314 publications

References 30 publications

Analytical Derivative Coupling for Multistate CASPT2 Theory

Analytical Derivative Coupling for Multistate CASPT2 Theory

Structure and mechanism of the reversible photoswitch of a fluorescent protein

Tuning the Excited-State Dynamics of GFP-Inspired Imidazolone Derivatives

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