Probing the Formation and Conformational Relaxation of Previtamin D and Analogues in Solution and in Lipid Bilayers.

The photosynthesis of vitamin D in mammalian skin results from UV-B irradiation of provitamin D (7-dehydrocholesterol, DHC) at ca. 290 nm. Upon return to the ground state, the hexatriene product, previtamin D, undergoes a conformational equilibration between helical gZg and more planar tZg and tZt forms.

Ultrafast excited state dynamics of provitamin D and analogs in solution and in lipid bilayers.

The photochemical ring-opening reaction of 7-dehydrocholesterol (DHC, provitamin D) is responsible for the light-initiated formation of vitamin D in mammalian skin membranes. Visible transient absorption spectroscopy was used to explore the excited state dynamics of DHC and two analogs: ergosterol (provitamin D) and DHC acetate free in solution and confined to lipid bilayers chosen to model the biological cell membrane. In solution, the excited state dynamics of the three compounds are nearly identical.

The Photoactive Excited State of the B-Based Photoreceptor CarH.

We have used transient absorption spectroscopy in the UV-visible and X-ray regions to characterize the excited state of CarH, a protein photoreceptor that uses a form of B, adenosylcobalamin (AdoCbl), to sense light. With visible excitation, a nanosecond-lifetime photoactive excited state is formed with unit quantum yield. The time-resolved X-ray absorption near edge structure difference spectrum of this state demonstrates that the excited state of AdoCbl in CarH undergoes only modest structural expansion around the central cobalt, a behavior similar to that observed for methylcobalamin rather than for AdoCbl free in solution.

Ultrafast Excited State Dynamics and Fluorescence from Vitamin B and Organometallic [Co]-C≡C-R Cobalamins.

Cobalamins are cobalt-centered cyclic tetrapyrrole ring-based molecules that provide cofactors for exceptional biological processes and possess unique and synthetically tunable photochemistry. Typical cobalamins are characterized by a visible absorption spectrum consisting of peaks labeled α, β, and sh. The physical basis of these peaks as having electronic origin or as a vibronic progression is ambiguous despite much investigation.

Exceptional Photochemical Stability of the Co-C Bond of Alkynyl Cobalamins, Potential Antivitamins B and Core Elements of B-Based Biological Vectors.

Alkynylcorrinoids are a class of organometallic B derivatives, recently rediscovered for use as antivitamins B and as core components of B-based biological vectors. They feature exceptional photochemical and thermal stability of their characteristic extra-short Co-C bond. We describe here the synthesis and structure of 3-hydroxypropynylcobalamin (HOPryCbl) and photochemical experiments with HOPryCbl, as well as of the related alkynylcobalamins: phenylethynylcobalamin and difluoro-phenylethynylcobalamin.

Ultrafast XANES Monitors Femtosecond Sequential Structural Evolution in Photoexcited Coenzyme B.

Polarized X-ray absorption near-edge structure (XANES) at the Co K-edge and broadband UV-vis transient absorption are used to monitor the sequential evolution of the excited-state structure of coenzyme B (adenosylcobalamin) over the first picosecond following excitation. The initial state is characterized by sub-100 fs sequential changes around the central cobalt. These are polarized first in the -direction orthogonal to the transition dipole and 50 fs later in the -direction along the transition dipole.

Probing the Excited State of Methylcobalamin Using Polarized Time-Resolved X-ray Absorption Spectroscopy.

We use picosecond time-resolved polarized X-ray absorption near-edge structure (XANES) measurements to probe the structure of the long-lived photoexcited state of methylcobalamin (MeCbl) and the cob(II)alamin photoproduct formed following photoexcitation of adenosylcobalamin (AdoCbl, coenzyme B). For MeCbl, we used 520 nm excitation and a time delay of 100 ps to avoid the formation of cob(II)alamin. We find only small spectral changes in the equatorial and axial directions, which we interpret as arising from small (<∼0.

Multispectral multidimensional spectrometer spanning the ultraviolet to the mid-infrared.

Multidimensional spectroscopy is the optical analog to nuclear magnetic resonance, probing dynamical processes with ultrafast time resolution. At optical frequencies, the technical challenges of multidimensional spectroscopy have hindered its progress until recently, where advances in laser sources and pulse-shaping have removed many obstacles to its implementation. Multidimensional spectroscopy in the visible and infrared (IR) regimes has already enabled respective advances in our understanding of photosynthesis and the structural rearrangements of liquid water.

Primed for Efficient Motion: Ultrafast Excited State Dynamics and Optical Manipulation of a Four Stage Rotary Molecular Motor.

All isomers of a four stage rotary molecular motor, dimethyl-tetrahydro-bi(cyclopenta[α]napthal-enylidene), are studied with ultrafast transient absorption spectroscopy. Single and two pulse excitations (pump and delayed repump with a different wavelength) are used to optically probe the excited state dynamics. These measurements demonstrate that this motor is not only designed for unidirectional isomerization, but is also "primed" for efficient rotary motion.

Electronic Interactions in the Bacterial Reaction Center Revealed by Two-Color 2D Electronic Spectroscopy.

The bacterial reaction center (BRC) serves as an important model system for understanding the charge separation processes in photosynthesis. Knowledge of the electronic structure of the BRC is critical for understanding its charge separation mechanism. While it is well-accepted that the "special pair" pigments are strongly coupled, the degree of coupling among other BRC pigments has been thought to be relatively weak.

Strongly coupled bacteriochlorin dyad studied using phase-modulated fluorescence-detected two-dimensional electronic spectroscopy.

Fluorescence-detected two-dimensional electronic spectroscopy (F-2DES) projects the third-order non-linear polarization in a system as an excited electronic state population which is incoherently detected as fluorescence. Multiple variants of F-2DES have been developed. Here, we report phase-modulated F-2DES measurements on a strongly coupled symmetric bacteriochlorin dyad, a relevant 'toy' model for photosynthetic energy and charge transfer.

Ultrafast X-ray Absorption Near Edge Structure Reveals Ballistic Excited State Structural Dynamics.

Polarized ultrafast time-resolved X-ray absorption near edge structure (XANES) allows characterization of excited state dynamics following excitation. Excitation of vitamin B, cyanocobalamin (CNCbl), in the αβ-band at 550 nm and the γ-band at 365 nm was used to uniquely resolve axial and equatorial contributions to the excited state dynamics. The structural evolution of the excited molecule is best described by a coherent ballistic trajectory on the excited state potential energy surface.

Primary processes in the bacterial reaction center probed by two-dimensional electronic spectroscopy.

In the initial steps of photosynthesis, reaction centers convert solar energy to stable charge-separated states with near-unity quantum efficiency. The reaction center from purple bacteria remains an important model system for probing the structure-function relationship and understanding mechanisms of photosynthetic charge separation. Here we perform 2D electronic spectroscopy (2DES) on bacterial reaction centers (BRCs) from two mutants of the purple bacterium , spanning the Q absorption bands of the BRC.

Stimulated Emission Enhancement Using Shaped Pulses.

Controlling stimulated emission is of importance because it competes with absorption and fluorescence under intense laser excitation. We performed resonant nonlinear optical spectroscopy measurements using femtosecond pulses shaped by π- or π/2-step phase functions and carried out calculations based on density matrix representation to elucidate the experimental results. In addition, we compared enhancements obtained when using other pulse shaping functions (chirp, third-order dispersion, and a time-delayed probe).

Controlling S2 Population in Cyanine Dyes Using Shaped Femtosecond Pulses.

Fast population transfer from higher to lower excited states occurs via internal conversion (IC) and is the basis of Kasha's rule, which states that spontaneous emission takes place from the lowest excited state of the same multiplicity. Photonic control over IC is of interest because it would allow direct influence over intramolecular nonradiative decay processes occurring in condensed phase. Here we tracked the S2 and S1 fluorescence yield for different cyanine dyes in solution as a function of linear chirp.

Investigating the role of human serum albumin protein pocket on the excited state dynamics of indocyanine green using shaped femtosecond laser pulses.

Differences in the excited state dynamics of molecules and photo-activated drugs either in solution or confined inside protein pockets or large biological macromolecules occur within the first few hundred femtoseconds. Shaped femtosecond laser pulses are used to probe the behavior of indocyanine green (ICG), the only Food and Drug Administration (FDA) approved near-infrared dye and photodynamic therapy agent, while free in solution and while confined inside the pocket of the human serum albumin (HSA) protein. Experimental findings indicate that the HSA pocket hinders torsional motion and thus mitigates the triplet state formation in ICG.

Polyatomic molecules under intense femtosecond laser irradiation.

Interaction of intense laser pulses with atoms and molecules is at the forefront of atomic, molecular, and optical physics. It is the gateway to powerful new tools that include above threshold ionization, high harmonic generation, electron diffraction, molecular tomography, and attosecond pulse generation. Intense laser pulses are ideal for probing and manipulating chemical bonding.

Solvent Environment Revealed by Positively Chirped Pulses.

The spectroscopy of large organic molecules and biomolecules in solution has been investigated using various time-resolved and frequency-resolved techniques. Of particular interest is the early response of the molecule and the solvent, which is difficult to study due to the ambiguity in assigning and differentiating inter- and intramolecular contributions to the electronic and vibrational populations and coherence. Our measurements compare the yield of fluorescence and stimulated emission for two laser dyes IR144 and IR125 as a function of chirp.

Solvation Stokes-Shift Dynamics Studied by Chirped Femtosecond Laser Pulses.

The early optical dynamic response, resulting population, and electronic coherence are investigated experimentally and modeled theoretically for IR144 in solution. The fluorescence and stimulated emission response are studied systematically as a function of chirp. The magnitude of the chirp effect on fluorescence and stimulated emission is found to depend quadratically on pulse energy, even where excitation probabilities range from 0.

Optical Response of Fluorescent Molecules Studied by Synthetic Femtosecond Laser Pulses.

The optical response of the fluorescent molecule IR144 in solution is probed by pairs of collinear pulses with intensity just above the linear dependence using two different pulse shaping methods. The first approach mimics a Michelson interferometer, while the second approach, known as multiple independent comb shaping (MICS), eliminates spectral interference. The comparison of interfering and non-interfering pulses reveals that linear interference between the pulses leads to the loss of experimental information at early delay times.