Ultrafast Fluorescence Spectroscopy Research Articles

Solvent-polarity dependence of ultrafast excited-state dynamics of trans-4-nitrostilbene.

Ultrafast excited-state dynamics of the simplest nitrostilbenes, namely trans-4-nitrostilbene (t-NSB), was studied in solvents of various polarities with ultrafast broadband time-resolved fluorescence and transient absorption spectroscopies, and by quantum-chemical computations. The results revealed that the initially excited S1(ππ*) state deactivation dynamics is strongly influenced by the solvent polarity. Specifically, the t-NSB S1-state lifetime decreases by three orders of magnitude from ∼60 ps in high-polarity solvents to ∼60 fs in nonpolar solvents. The strong solvent-polarity dependence arises from the differences in dipole moments among the S1 and relevant states, including the major intersystem crossing (ISC) receiver triplet states, and therefore, the solvent polarity can modulate their relative energies and ISC rates. In nonpolar solvents, the sub-100 fs lifetime is due to a combination of efficient ISC and internal conversion. In medium-polarity solvents, the S1-state population decays via a competing ISC relaxation mechanism in a biphasic manner, and the ISC rates are found to obey the inverse energy gap law of the strong coupling case. In high-polarity solvents, the S1 state is stabilized to a much lower energy such that ISC becomes energetically infeasible, and the S1 state decays via barrier crossing along the torsion angle of the central ethylenic bond to the nonfluorescent perpendicular configuration. Regardless of the initial S1-state deactivation pathways in various solvents, the excited-state population is ultimately trapped in the metastable T1-state perpendicular configuration, at which a slower ISC occurs to bring the system to the ground state and bifurcate into either trans or cis form of NSB.

Cryo-EM and femtosecond spectroscopic studies provide mechanistic insight into the energy transfer in CpcL-phycobilisomes

Phycobilisomes (PBS) are the major light harvesting complexes of photosynthesis in the cyanobacteria and red algae. CpcL-PBS is a type of small PBS in cyanobacteria that transfers energy directly to photosystem I without the core structure. Here we report the cryo-EM structure of the CpcL-PBS from the cyanobacterium Synechocystis sp. PCC 6803 at 2.6-Å resolution. The structure shows the CpcD domain of ferredoxin: NADP+ oxidoreductase is located at the distal end of CpcL-PBS, responsible for its attachment to PBS. With the evidence of ultrafast transient absorption and fluorescence spectroscopy, the roles of individual bilins in energy transfer are revealed. The bilin 1Iβ822 located near photosystem I has an enhanced planarity and is the red-bilin responsible for the direct energy transfer to photosystem I.

Fluorescence Resonance Energy Transfer Properties and Auger Recombination Suppression in Supraparticles Self-Assembled from Colloidal Quantum Dots

Colloid quantum dots (CQDs) are recognized as an ideal material for applications in next-generation optoelectronic devices, owing to their unique structures, outstanding optical properties, and low-cost preparation processes. However, monodisperse CQDs cannot meet the requirements of stability and collective properties for device applications. Therefore, it is urgent to build stable 3D multiparticle systems with collective physical and optical properties, which is still a great challenge for nanoscience. Herein, we developed a modified microemulsion template method to synthesize quantum dot supraparticles (QD-SPs) with regular shapes and a high packing density, which is an excellent research platform for ultrafast optical properties of composite systems. The redshift of the steady-state fluorescence spectra of QD-SPs compared to CQD solutions indicates that fluorescence resonance energy transfer (FRET) occurred between the CQDs. Moreover, we investigated the dynamic processes of energy transfer in QD-SPs by time-resolved ultrafast fluorescence spectroscopy. The dynamic redshift and lifetime changes of the spectra further verified the existence of rapid energy transfer between CQDs with different exciton energies. In addition, compared with CQD solutions, the steady-state fluorescence lifetime of SPs increased and the fluorescence intensity decreased slowly with increasing temperature, which indicates that the SP structure suppressed the Auger recombination of CQDs. Our results provide a practical approach to enhance the coupling and luminescence stability of CQDs, which may enable new physical phenomena and improve the performance of optoelectronic devices.

Sequence-specific binding behavior of coralyne toward triplex DNA: An ultrafast time-resolved fluorescence spectroscopy study.

Triplex DNA structure has potential therapeutic application in inhibiting the expression of genes involved in cancer and other diseases. As a DNA-targeting antitumor and antibiotic drug, coralyne shows a remarkable binding propensity to triplex over canonical duplex and thus can modulate the stability of triplex structure, providing a prospective gene targeting strategy. Much less is known, however, about coralyne-binding interactions with triplex. By combining multiple steady-state spectroscopy with ultrafast fluorescence spectroscopy, we have investigated the binding behaviors of coralyne with typical triplexes. Upon binding with a G-containing triplex, the fluorescence of coralyne is markedly quenched owing to the photoinduced electron transfer (PET) of coralyne with the G base. Systematic studies show that the PET rates are sensitive to the binding configuration and local microenvironment, from which the coexisting binding modes of monomeric (full and partial) intercalation and aggregate stacking along the sugar-phosphate backbone are distinguished and their respective contributions are determined. It shows that coralyne has preferences for monomeric intercalation within CGG triplex and pure TAT triplex, whereas CGC+ triplex adopts mainly backbone binding of coralyne aggregates due to charge repulsion, revealing the sequence-specific binding selectivity. The triplex-DNA-induced aggregation of coralyne could be used as a probe for recognizing the water content in local DNA structures. The strong π-π stacking of intercalated coralyne monomer with base-triplets plays an important role in stabilizing the triplex structure. These results provide mechanistic insights for understanding the remarkable propensity of coralyne in selective binding to triplex DNA and shed light on the prospective applications of coralyne-triplex targeted anti-gene therapeutics.

Excitation relaxation dynamics of carotenoids constituting the diadinoxanthin cycle.

Carotenoids (Cars) exhibit two functions in photosynthesis, light-harvesting and photoprotective functions, which are performed through the excited states of Cars. Therefore, increasing our knowledge on excitation relaxation dynamics of Cars is important for understanding of the functions of Cars. In light-harvesting complexes, there exist Cars functioning by converting the π-conjugation number in response to light conditions. It is well known that some microalgae have a mechanism controlling the conjugation number of Cars, called as the diadinoxanthin cycle; diadinoxanthin (10 conjugations) is accumulated under low light, whereas diatoxanthin (11 conjugations) appears under high light. However, the excitation relaxation dynamics of these two Cars have not been clarified. In the present study, we investigated excitation relaxation dynamics of diadinoxanthin and diatoxanthin in relation to their functions, by the ultrafast fluorescence spectroscopy. After an excitation to the S2 state, the intramolecular vibrational redistribution occurs, followed by the internal conversion to the S1 state. The S2 lifetimes were analyzed to be 175fs, 155fs, and 140fs in diethyl ether, ethanol, and acetone, respectively, for diadinoxanthin, and 155fs, 135fs, and 125fs in diethyl ether, ethanol, and acetone, respectively for diatoxanthin. By converting diadinoxanthin to diatoxanthin, the absorption spectra shift to longer wavelengths by 5-7nm, and lifetimes of S2 and S1 states decrease by 11-13% and 52%, respectively. Differences in levels and lifetimes of excited states between diadinoxanthin and diatoxanthin are small; therefore, it is suggested that changes in the energy level of chlorophyll a are necessary to efficiently control the functions of the diadinoxanthin cycle.

Ultrafast photooxidation of protein-bound anionic flavin radicals

The photophysical properties of anionic semireduced flavin radicals are largely unknown despite their importance in numerous biochemical reactions. Here, we studied the photoproducts of these intrinsically unstable species in five different flavoprotein oxidases where they can be stabilized, including the well-characterized glucose oxidase. Using ultrafast absorption and fluorescence spectroscopy, we unexpectedly found that photoexcitation systematically results in the oxidation of protein-bound anionic flavin radicals on a time scale of less than ∼100 fs. The thus generated photoproducts decay back in the remarkably narrow 10- to 20-ps time range. Based on molecular dynamics and quantum mechanics computations, positively charged active-site histidine and arginine residues are proposed to be the electron acceptor candidates. Altogether, we established that, in addition to the commonly known and extensively studied photoreduction of oxidized flavins in flavoproteins, the reverse process (i.e., the photooxidation of anionic flavin radicals) can also occur. We propose that this process may constitute an excited-state deactivation pathway for protein-bound anionic flavin radicals in general. This hitherto undocumented photochemical reaction in flavoproteins further extends the family of flavin photocycles.

Synergistic dynamics of photoionization and photoinduced electron transfer probed by laser flash photolysis and ultrafast fluorescence spectroscopy.

Photoionization (PI) and photoinduced electron transfer (PET) dynamics of coumarin 450 (C450) in micelles were investigated in the time domains of micro to femtoseconds using steady-state and time-resolved absorption and fluorescence spectroscopy. The PI of C450 occurs inside the micelles leads to the formation of C450 cation radical (CR) and hydrated electron, which is characterized by the respective transient absorption. The PI of C450 is monophotonic in nature and the yield is dependent on the charge of the micelles. The observation of amine CR in the transient absorption confirms the PET from amine to the excited state of C450 in micelles, which results in the quenching of both fluorescence intensity and lifetime. The decrease in femtosecond fluorescent decay of C450 and the absence of transient C450 radical anion in the presence of amine implies that the concerted ultrafast PET promoted PI and PET to the C450 CR-electron pair. The decrease in the time constant for the formation of relaxed state in the presence of amines is due to the ultrafast PET to the C450 CR-electron pair, which prevents the formation of a relaxed state through recombination at a longer time scale. In the present investigation, the recombination dynamics of the CR-electron pair is justified as one of the origins of the slow solvation in micelles. The influence of amine concentration on the decay of C450 CR indicates ET reaction between C450 CR and amine, which is further confirmed by the bleach recovery of C450 ground state in the presence of amine.

Sulfated-β-cyclodextrin templated aggregation of a metachromatic dye, Basic Orange 21: A photophysical investigation

ABSTRACT The recently discovered molecular rotor and supravital dye, Basic Orange-21 (BO21), has shown improved differential counting of leukocytes through metachromasia (change in colour), which is believed to be due to the aggregation of chromophores. Thus, investigation on the aggregation behaviour of BO21 holds special importance and may help us to delineate its spectroscopic behaviour. In this contribution, we have investigated the photophysical behaviour of BO21 in presence of a negatively charged supramolecular host, sulfated-β-cyclodextrin (SC), using ground-state absorption, steady-state and time-resolved emission spectroscopy including ultrafast fluorescence spectroscopy, and molecular docking calculations. The spectral characteristics of BO21 display extreme modulations in presence of SCD, with features similar to that observed for BO21 during its metachromatic behaviour. These spectroscopic features have been rationalised in terms of aggregation of BO21 molecules in the vicinity of SCD, which are also extremely sensitive to external stimuli, such as, ionic strength and temperature of the medium.

Fluorescent iron‑sulfur centers: Photochemistry of the PetA Rieske protein from Aquifex aeolicus

Cytochrome bc1 complexes are energy-transducing enzymes and key components of respiratory electron chains. They contain Rieske 2Fe2S proteins that absorb very weakly in the visible absorption region compared to the heme cofactors of the cytochromes, but are known to yield photoproducts. Here, the photoreactions of isolated Rieske proteins from the hyperthermophilic bacterium Aquifex aeolicus are studied in two redox states using ultrafast transient fluorescence and absorption spectroscopy. We provide evidence, for the first time in iron‑sulfur proteins, of very weak fluorescence of the excited state, in the oxidized as well as the reduced state. The excited states of the oxidized and reduced forms decay in 1.5 ps and 30 ps, respectively. In both cases they give rise to product states with lifetimes beyond 1 ns, reflecting photo-reduction of oxidized centers as well as photo-oxidation of reduced centers. Potential reaction partners are discussed and studied using site-directed mutagenesis. For the reduced state, a nearby disulfide bridge is suggested as an electron acceptor. The resulting photoproducts in either state may play a role in photoactivation processes.

Ultrafast Fluorescence Spectroscopy via Upconversion and Its Applications in Biophysics.

In this review, the experimental set-up and functional characteristics of single-wavelength and broad-band femtosecond upconversion spectrophotofluorometers developed in our laboratory are described. We discuss applications of this technique to biophysical problems, such as ultrafast fluorescence quenching and solvation dynamics of tryptophan, peptides, proteins, reduced nicotinamide adenine dinucleotide (NADH), and nucleic acids. In the tryptophan dynamics field, especially for proteins, two types of solvation dynamics on different time scales have been well explored: ~1 ps for bulk water, and tens of picoseconds for “biological water”, a term that combines effects of water and macromolecule dynamics. In addition, some proteins also show quasi-static self-quenching (QSSQ) phenomena. Interestingly, in our more recent work, we also find that similar mixtures of quenching and solvation dynamics occur for the metabolic cofactor NADH. In this review, we add a brief overview of the emerging development of fluorescent RNA aptamers and their potential application to live cell imaging, while noting how ultrafast measurement may speed their optimization.

Long living excited state of protonated adenosine unveiled by ultrafast fluorescence spectroscopy and density functional theoretical study.

Adenosine (Ado) possesses ultrafast nonradiative dynamics accounting for its remarkably high photostability. The deactivation dynamics of Ado after protonation in an aqueous solution remains an elusive issue. Herein we report an investigation of the excited state dynamics of protonated Ado (AdoH+) performed using ultrafast time-resolved fluorescence spectroscopy combined with density functional theoretical calculation. The result obtained from comparison of conformers with protonation at different sites revealed that the syn-conformer with protonation occurring at the N3 position (syn-N3) is the predominant form of AdoH+ in the ground state, similar to that of Ado. In contrast, the fluorescence of AdoH+ with maximum intensity at 385 nm, significantly red-shifted from that of Ado, displaying decay dynamics composed of an ultrafast component with the lifetime of ∼0.5 ps and a slower one of ∼2.9 ns. The former is because of the decay of the syn-N3 conformer, similar to that reported for AdoH+ under the gas phase condition. The latter is due to the syn-N1 conformer formed via ultrafast proton transfer of the syn-N3. The excited state of syn-N1 has a peculiar nonplanar conformation over the purine molecule, which is responsible for the substantial Stokes shift showed in the fluorescence spectrum and correlates with a large energy barrier for nonradiative decay likely involving a reversed proton transfer. This study demonstrates the importance of protonation and solvent environment in altering dramatically the excited states of Ado, providing insight for better understanding nonradiative dynamics of both the monomeric bases and the oligomeric or polymeric DNAs.

Altered relaxation dynamics of excited state reactions by confinement in reverse micelles probed by ultrafast fluorescence up-conversion.

Chemical reactions in confined environments are important in areas as diverse as heterogenous catalysis, environmental chemistry and biochemistry, yet they are much less well understood than the equivalent reactions in either the gas phase or in free solution. The understanding of chemical reactions in solution was greatly enhanced by real time studies of model reactions, through ultrafast spectroscopy (especially when supported by molecular dynamics simulation). Here we review some of the efforts that have been made to adapt this approach to the investigation of reactions in confined media. Specifically, we review the application of ultrafast fluorescence spectroscopy to measure reaction dynamics in the nanoconfined water phase of reverse micelles, as a function of the droplet radius and the charge on the interface. Methods of measurement and modelling of the reactions are outlined. In all of the cases studied (which are focused on ultrafast intramolecular reactions) the effect of confinement was to suppress the reaction. Even in the largest micelles the result in the bulk aqueous phase was not usually recovered, suggesting an important role for specific interactions between reactant and environment, for example at the interface. There was no simple one-to-one correspondence with direct measures of the dynamics of the confined phase. Thus, understanding the effect of confinement on reaction rate appears to require not only knowledge of the dynamics of the reaction in solutions and the effect of confinement on the medium, but also of the interaction between reactant and confining medium.

A leap in quantum efficiency through light harvesting in photoreceptor UVR8

Plants utilize a UV-B (280 to 315 nm) photoreceptor UVR8 (UV RESISTANCE LOCUS 8) to sense environmental UV levels and regulate gene expression to avoid harmful UV effects. Uniquely, UVR8 uses intrinsic tryptophan for UV-B perception with a homodimer structure containing 26 structural tryptophan residues. However, besides 8 tryptophans at the dimer interface to form two critical pyramid perception centers, the other 18 tryptophans’ functional role is unknown. Here, using ultrafast fluorescence spectroscopy, computational methods and extensive mutations, we find that all 18 tryptophans form light-harvesting networks and funnel their excitation energy to the pyramid centers to enhance light-perception efficiency. We determine the timescales of all elementary tryptophan-to-tryptophan energy-transfer steps in picoseconds to nanoseconds, in excellent agreement with quantum computational calculations, and finally reveal a significant leap in light-perception quantum efficiency from 35% to 73%. This photoreceptor is the first system discovered so far, to be best of our knowledge, using natural amino-acid tryptophans to form networks for both light harvesting and light perception.

Energy transfer dynamics in B-phycoerythrin from the red alga Porphyridium purpureum

The B-phycoerythrin hexamer (αβ)6γ of Porphyridium purpureum was isolated and purified. The absorption, circular dichroism, fluorescence and ultrafast time-resolved spectra were obtained. The results showed a double absorption peak at 545 nm and 565 nm and a shoulder peak at 498 nm, and fluorescence emission maxima at 580 nm and 620 nm were observed. The circular dichroism spectra in the near-ultraviolet region were obtained and resolved for the first time, which showed that the two peaks at 260 nm and 305 nm were considered to be correlated to phenylalanine (Phe) and tryptophan (Trp) in a conservative hydrophobic microenvironment, respectively. The circular dichroism spectra in the visible region showed that PEB139α/PEB158β and PEB82α/PEB82β existed as two exciton-coupled bilin pairs. Energy transfer within the exciton-coupled pairs was by exciton splitting, while between the exciton-coupled pairs was by Förster resonance. From the studies of the energy transfer dynamics by ultrafast time-resolved fluorescence spectroscopy, it was confirmed that the energy transfer of the B-PE hexamer had three time components of 8 ps, 60 ps, and 1200 ps. In addition, the internal energy transfer pathways of B-phycoerythrin hexamer were identified by deconvoluting the fluorescence decay curve at different detection wavelengths.

Mapping the excitation energy migration pathways in phycobilisomes from the cyanobacterium Acaryochloris marina

In this study, we use ultrafast time-resolved absorption and fluorescence spectroscopies to examine A. marina phycobilisomes isolated from cells grown under light of different intensities and spectral regimes. Investigations were performed at room temperature and at 77 K. The study demonstrates that if complexes are stabilized by high phosphate (900 mM) buffer, there are no differences between them in temporal and spectral properties of fluorescence. However, when the complexes are allowed to disassemble into trimers in low phosphate (50 mM) buffer, differences are clearly observed. The fluorescence properties of intact or disassembled phycobilisomes from cells grown in low intensity white light are unresponsive to variation in phosphate concentration. This antenna complex was further studied in detail with application of femtosecond time-resolved absorption at room temperature. Combined spectroscopic and kinetic analysis of time-resolved fluorescence and absorption data of this antenna allowed us to identify spectrally different forms of phycocyanobilins and to propose a simplified model of how they could be distributed within the phycobilisome structure.

Using Bulk-like Nanocrystals To Probe Intrinsic Optical Gain Characteristics of Inorganic Lead Halide Perovskites.

Following the introduction of perovskites for photovoltaic solar energy conversion, the use of these materials as a general purpose optoelectronic material for displays, lighting, and lasing has been explored. However, while reports on stimulated emission and lasing by perovskites show great promise, a comprehensive quantification of their optical gain characteristics is lacking. Here, we measure gain coefficients, clarify the gain mechanism, and explore the gain dynamics of colloidal CsPbBr3 nanocrystals by deploying a unique combination of broadband transient absorption and ultrafast fluorescence spectroscopy. Opposite from current literature, we show that optical gain in such nanocrystals is supported by stimulated emission from free carriers, and not from excitons or biexcitons. Importantly, we demonstrate that the concomitant gain coefficients and thresholds agree with literature results reported for perovksite thin films. Finally, we show that, even in the case of fully inorganic lead halide perovskites, a cooling bottleneck hampers the development of net stimulated emission at high excitation density. Based on these results, we propose that bulk-like colloidal nanocrystals in general offer a unique testbed to quantify optical gain of novel photonic materials and in particular for lead halide perovskites.

Ultrafast dynamics-driven biomolecular recognition where fast activities dictate slow events

In general, biological macromolecules require significant dynamical freedom to carry out their different functions, including signal transduction, metabolism, catalysis and gene regulation. Effectors (ligands, DNA and external milieu, etc) are considered to function in a purely dynamical manner by selectively stabilizing a specific dynamical state, thereby regulating biological function. In particular, proteins in presence of these effectors can exist in several dynamical states with distinct binding or enzymatic activity. Here, we have reviewed the efficacy of ultrafast fluorescence spectroscopy to monitor the dynamical flexibility of various proteins in presence of different effectors leading to their biological activity. Recent studies demonstrate the potency of a combined approach involving picosecond-resolved Forster resonance energy transfer, polarisation-gated fluorescence and time-dependent stokes shift for the exploration of ultrafast dynamics in biomolecular recognition of various protein molecules. The allosteric protein-protein recognition following differential protein-DNA interaction is shown to be a consequence of some ultrafast segmental motions at the C-terminal of Gal repressor protein dimer with DNA operator sequences OE and OI. Differential ultrafast dynamics at the C-terminal of λ-repressor protein with two different operator DNA sequences for the protein-protein interaction with different strengths is also reviewed. We have also systemically briefed the study on the role of ultrafast dynamics of water molecules on the functionality of enzyme proteins alpha-chymotrypsin and deoxyribonuclease I. The studies on the essential ultrafast dynamics at the active site of the enzyme alpha-chymotrypsin by using an anthraniloyl fluorescent extrinsic probe covalently attached to the serine-195 residue for the enzymatic activity at homeothermic condition has also been reviewed. Finally, we have highlighted the evidence that a photoinduced dynamical event dictates the molecular recognition of a photochromic ligand, dihydroindolizine with the serine protease alpha-chymotrypsin and with a liposome (L-a-phosphatidylcholine).

Ultrafast Through-Space Electronic Energy Transfer in Molecular Dyads Built around Dynamic Spacer Units.

A pair of complementary molecular dyads have been synthesized around a 1,2-diaminocyclohexyl spacer that itself undergoes ring inversion. Despite these conformational exchange processes, the donor and acceptor occupy quite restricted spatial regions, and they are not interchangeable. The donor and acceptor pair comprise disparate boron dipyrromethene dyes selected to display favorable electronic energy transfer (EET). Steady-state emission spectroscopy confirms that through-space EET from donor to acceptor is almost quantitative, aided by the relatively short separations. Ultrafast time-resolved fluorescence spectroscopy has allowed determination of the rates of EET for both dyads. Surprisingly, in view of the close proximity of donor and acceptor (center-to-center separations less than 20 Å), the EET dynamics are well-accounted for in terms of the computed molecular conformations and conventional Förster theory. One dyad appears as a single family of conformations, but EET for the second dyad corresponds to dual-exponential kinetics. In this latter case, an intramolecular hydrogen bond helps stabilize an open geometry, wherein EET is relatively slow.

The energy transfer dynamics in C-phycocyanin hexamer from <italic>Spirulina platensis</italic>

Photosynthesis is an ancient and important conversion of solar light to biological energy in photosynthetic organisms. Phycobilisome (PBS) system in cyanobacteria and red algae is one of two major light-harvesting systems of photosynthetic oxygen organisms. PBS is aggregation of water-soluble phycobiliproteins (PBPs) which are distinctively colored group of disk-shaped macromolecular proteins bearing covalently attached open-chain tetrapyrroles known as phycobilins. PBPs are capable to absorb solar light and transfer energy to the chlorophyll of photosynthetic reaction center with a high efficiency. Light energy absorbed by phycobilins is transmitted between the subunits, and then transmitted between different PBPs, finally to the photoreaction center located on the thylakoid membrane. The energy transfer between PBPs is very fast and high efficiency. Because energy can be transferred in different chromophores and there are a large number of chromophores in PBPs, the energy transfer path of PBPs is complex and diverse. In the past 20 years, energy transfer process of PBPs have been analyzed by time-resolved fluorescence spectroscopy because a large shift of the fluorescence peaks can be clearly observed. For example, Zhang et al. estimated the time constant for energy transfer from PC (rod) to APC (core) as 18 ps and 55 ps based on PBPs crystal structures and time-resolved fluorescence spectra of the rod-core complex. However, due to the technical bottleneck of theoretical simulation and spectral experimental detection, the research on the physical mechanism of photosynthetic energy transfer is relatively slow. In recent years, the ultrafast spectrum time resolution has been continuously improved, which provides the conditions for studying the energy transfer of PBPs. In this paper, the mechanism of energy transfer in C-phycocyanin hexamer from Spirulina platensis was studied by ultrafast time-resolved fluorescence spectroscopy. The energy transfer pathway and the related transfer parameters in the C-phycocyanin hexamer were identified by the fitting of the fluorescence decay curve at different detection wavelengths. It was proved that the energy transfer of C-phycocyanin hexamer had four time components from the experiment, which are 6, 22, 280 and 1470 ps, respectively, then the time components were identified and discussed. This study reflects the energy transfer kinetics between different chromophores in C-phycocyanin and provides important experimental data for solving the relationship between the structure and function of PBPs. At the same time, our study has important value for the understanding of the primary process of photosynthesis. In the future, if the resolution of a streak camera can be raised to femtoseconds, it is possible to find more energy transfer paths, and thus it is expected to make a breakthrough in the ultrafast and ultra-efficient energy transfer mechanism of PBPs.

Prying into Hydration Water in Amyloidogenic Intrinsically Disordered Proteins

The hydration water layer, termed as ‘biological water’, present at the protein-water interface shares an intimate relationship with protein functions. However, little is known about hydration water in intrinsically disordered proteins (IDPs) that are capable of adopting a multitude of conformational states. Recent studies have revealed that many IDPs, due to their astonishing conformational plasticity, can readily convert into highly ordered amyloid aggregates that are associated with several deadly human diseases. For instance, α-synuclein is an IDP present in nerve cells and aggregates into amyloids via toxic oligomeric intermediates in Parkinson's disease. To address the role of water in the pathological conversion of α-synuclein, we utilized a combination of ultrafast measurements and all-atom molecular dynamics simulations. We created a number of single cysteine mutants and labeled these variants using environment-sensitive thiol-active fluorophore. We then monitored the solvation dynamics using ultrafast fluorescence spectroscopy that is capable of capturing the femtosecond time-resolved snapshots of water molecules around the polypeptide chain. These site-specific ultrafast studies allowed us to construct the hydration map of α-synuclein. Our studies revealed the presence of three distinct types of water: bulk-, hydration- and confined water. Atomistic molecular dynamics simulations supported our experimental observations and provided insights into the molecular origin of confined water. Our results from this experimental-simulation approach reveal trapping of interfacial water near the amyloidogenic hotspot that triggers the conversion of IDPs into pathological amyloids via entropically favorable water exclusion events during disorder-to-order amyloid transition. Our findings provide an experimental underpinning of significant water rearrangements associated with both chain desolvation and water confinement upon amyloid formation from IDPs. I will also discuss our new results on the hydration dynamics of a diverse range of amyloidogenic IDPs possessing different conformational preferences due to varied chain-chain and chain-solvent interactions.