Franklin D. Fuller

Vibronic coherence in oxygenic photosynthesis

Photosynthesis powers life on our planet. The basic photosynthetic architecture consists of antenna complexes that harvest solar energy and reaction centres that convert the energy into stable separated charge. In oxygenic photosynthesis, the initial charge separation occurs in the photosystem II reaction centre, the only known natural enzyme that uses solar energy to split water. Both energy transfer and charge separation in photosynthesis are rapid events with high quantum efficiencies. In recent nonlinear spectroscopic experiments, long-lived coherences have been observed in photosynthetic antenna complexes, and theoretical work suggests that they reflect underlying electronic-vibrational resonances, which may play a functional role in enhancing energy transfer. Here, we report the observation of coherent dynamics persisting on a picosecond timescale at 77 K in the photosystem II reaction centre using two-dimensional electronic spectroscopy. Supporting simulations suggest that the coherences are of a mixed electronic-vibrational (vibronic) nature and may enhance the rate of charge separation in oxygenic photosynthesis.

Conformational dynamics of single pre-mRNA molecules during in vitro splicing

The spliceosome is a complex small nuclear RNA (snRNA)-protein machine that removes introns from pre-mRNAs via two successive phosphoryl transfer reactions. The chemical steps are isoenergetic, yet splicing requires at least eight RNA-dependent ATPases responsible for substantial conformational rearrangements. To comprehensively monitor pre-mRNA conformational dynamics, we developed a strategy for single-molecule FRET (smFRET) that uses a small, efficiently spliced yeast pre-mRNA, Ubc4, in which donor and acceptor fluorophores are placed in the exons adjacent to the 5′ and 3′ splice sites. During splicing in vitro, we observed a multitude of generally reversible time- and ATP-dependent conformational transitions of individual pre-mRNAs. The conformational dynamics of branchpoint and 3′–splice site mutants differ from one another and from wild type. Because all transitions are reversible, spliceosome assembly appears to be occurring close to thermal equilibrium.

Experimental implementations of two-dimensional fourier transform electronic spectroscopy.

Two-dimensional electronic spectroscopy (2DES) reveals connections between an optical excitation at a given frequency and the signals it creates over a wide range of frequencies. These connections, manifested as cross-peak locations and their lineshapes, reflect the underlying electronic and vibrational structure of the system under study. How these spectroscopic signatures evolve in time reveals the system dynamics and provides a detailed picture of coherent and incoherent processes. 2DES is rapidly maturing and has already found numerous applications, including studies of photosynthetic energy transfer and photochemical reactions and many-body interactions in nanostructured materials. Many systems of interest contain electronic transitions spanning the ultraviolet to the near infrared and beyond. Most 2DES measurements to date have explored a relatively small frequency range. We discuss the challenges of implementing 2DES and compare and contrast different approaches in terms of their information content, ease of implementation, and potential for broadband measurements.

Tight-binding model of the photosystem II reaction center: application to two-dimensional electronic spectroscopy

We propose an optimized tight-binding electron-hole model of the photosystem II (PSII) reaction center (RC). Our model incorporates two charge separation pathways and spatial correlations of both static disorder and fast fluctuations of energy levels. It captures the main experimental features observed in time-resolved two-dimensional (2D) optical spectra at 77K: peak pattern, lineshapes and time traces. Analysis of 2D spectra kinetics reveals that specific regions of the 2D spectra of the PSII RC are sensitive to the charge transfer states. We find that the energy disorder of two peripheral chlorophylls is four times larger than the other RC pigments.

Effects of chirp on two-dimensional Fourier transform electronic spectra

We examine the effect that pulse chirp has on the shape of two- dimensional electronic spectra through calculations and experiments. For the calculations we use a model two electronic level system with a solvent interaction represented by a simple Gaussian correlation function and compare the resulting spectra to experiments carried out on an organic dye molecule (Rhodamine 800). Both calculations and experiments show that distortions due to chirp are most significant when the pulses used in the experiment have different amounts of chirp, introducing peak shape asymmetry that could be interpreted as spectrally dependent relaxation. When all pulses have similar chirp the distortions are reduced but still affect the anti-diagonal symmetry of the peak shapes and introduce negative features that could be interpreted as excited state absorption.

Simulations of the Two-Dimensional Electronic Spectroscopy of the Photosystem II Reaction Center

We report simulations of the two-dimensional electronic spectroscopy of the Q(y) band of the D1-D2-Cyt b559 photosystem II reaction center at 77 K. We base the simulations on an existing Hamiltonian that was derived by simultaneous fitting to a wide range of linear spectroscopic measurements and described within modified Redfield theory. The model obtains reasonable agreement with most aspects of the two-dimensional spectra, including the overall peak shapes and excited state absorption features. It does not reproduce the rapid equilibration from high energy to low energy excitonic states evident by a strong cross-peak below the diagonal. We explore modifications to the model to incorporate new structural data and improve agreement with the two-dimensional spectra. We find that strengthening the system-bath coupling and lowering the degree of disorder significantly improves agreement with the cross-peak feature, while lessening agreement with the relative diagonal/antidiagonal width of the 2D spectra. We conclude that two-dimensional electronic spectroscopy provides a sensitive test of excitonic models of the photosystem II reaction center and discuss avenues for further refinement of such models.

Toward Broad Bandwidth 2-D Electronic Spectroscopy: Correction of Chirp From a Continuum Probe

Recent implementations of 2-D spectroscopy in the pump-probe geometry using a pulse-shaper demonstrate the ease with which frequency-resolved pump-probe experiments can be readily adapted to 2-D methods. Many frequency-resolved pump-probe experiments employ continuum probes to observe a broad range of electronic transitions. These continuum probes are often chirped, leading to distortions that can be post corrected by characterizing the chirp and appropriately adjusting the observed wavelength-dependent pump-probe time delay. We present an analogous chirp-correction scheme for 2-D spectroscopy, facilitating the use of continuum probing to expand the frequency information available in 2-D spectroscopy experiments. We demonstrate the method through experiments and simulations of a laser dye in solution.

Fast second-harmonic generation frequency-resolved optical gating using only a pulse shaper.

In many ultrafast contexts, a collinear pulse-shaping frequency-resolved optical gating (FROG) technique is desired. Some applicable techniques already exist, but they suffer from one of two issues: either they require many time points to allow for Fourier filtering, or they do not yield a traditional FROG trace. To overcome these issues, we propose and demonstrate a fast new phase-cycled FROG technique using a pulse shaper.

Two-dimensional electronic spectroscopy signatures of the glass transition

Two-dimensional electronic spectroscopy is a sensitive probe of solvation dynamics. Using a pump-probe geometry with a pulse shaper (Optics Express 15 (2007), 16681-16689; Optics Express 16 (2008), 17420-17428), we present temperature dependent 2D spectra of laser dyes dissolved in glass-forming solvents. At low waiting times, the system has not yet relaxed, resulting in a spectrum that is elongated along the diagonal. At longer times, the system loses its memory of the initial excitation frequency, and the 2D spectrum rounds out. As the temperature is lowered, the time scale of this relaxation grows, and the elongation persists for longer waiting times. This can be measured in the ratio of the diagonal width to the anti-diagonal width; the behavior of this ratio is representative of the frequency-frequency correlation function (Optics Letters 31 (2006), 3354-3356). Near the glass transition temperature, the relaxation behavior changes. Understanding this change is important for interpreting temperature-dependent dynamics of biological systems.

Two-Dimensional Electronic Spectroscopy of the Q y Band of Photosystem II Reaction Centers

We present two-dimensional electronic spectroscopy studies on the dynamics of D1-D2 cyt.b559 reaction center complexes from plant photosystem II at 77 K. Our two-dimensional spectra are compared with models based on current theory.