Light-induced Radical Research Articles

Bio-inspired Photocatalytic Ruthenium Complexes: Synthesis, Optical Properties, and Solvatochromic Effect.

We report the synthesis, characterization, and photo-physical properties of two new rutheniumII -phenol-imidazole complexes. These bio-mimetic complexes have potential as photocatalysts for water splitting. Owing to their multiple phenol-imidazole groups, they have a higher probability of light-induced radical formation than existing complexes. The newly synthesized complexes show improved overlap with the solar spectrum compared to other rutheniumII -phenol-imidazole complexes, and their measured lifetimes are suitable for light-induced radical formation. In addition, we conducted solvatochromic absorption measurements, which elegantly follow Marcus theory, and demonstrate the symmetry differences between the two complexes. The solvatochromic measurements further imply electron localization onto one of the ligands. The new complexes may find applications in photocatalysis, dye-sensitized solar cells, biomedicine, and sensing. Moreover, their multiple chelating units make them promising candidates for light-activated metal organic radical frameworks, i.e. metal-organic frameworks that contain organic radicals activated by light.

Radical Difluororomethylation of Thiols with Difluoromethylphosphonium Triflate under Photoredox Catalysis.

A convenient, visible light-induced radical difluoromethylation of aryl-, heteroaryl-, and alkylthiols with difluoromethyltriphenylphosphonium triflate was developed to afford various difluoromethyl thioethers in moderate to excellent yields. The key reaction features include the use of a readily available CF2H radical source, mild reaction conditions, and excellent chemoselective thiol-difluoromethylation.

Light-Induced Radical Formation and Isomerization of an Aromatic Thiol in Solution Followed by Time-Resolved X-ray Absorption Spectroscopy at the Sulfur K-Edge

We applied time-resolved sulfur-1s absorption spectroscopy to a model aromatic thiol system as a promising method for tracking chemical reactions in solution. Sulfur-1s absorption spectroscopy allows tracking multiple sulfur species with a time resolution of ∼70 ps at synchrotron radiation facilities. Experimental transient spectra combined with high-level electronic structure theory allow identification of a radical and two thione isomers, which are generated upon illumination with 267 nm radiation. Moreover, the regioselectivity of the thione isomerization is explained by the resulting radical frontier orbitals. This work demonstrates the usefulness and potential of time-resolved sulfur-1s absorption spectroscopy for tracking multiple chemical reaction pathways and transient products of sulfur-containing molecules in solution.

Inhomogeneous ensembles of radical pairs in chemical compasses.

The biophysical basis for the ability of animals to detect the geomagnetic field and to use it for finding directions remains a mystery of sensory biology. One much debated hypothesis suggests that an ensemble of specialized light-induced radical pair reactions can provide the primary signal for a magnetic compass sensor. The question arises what features of such a radical pair ensemble could be optimized by evolution so as to improve the detection of the direction of weak magnetic fields. Here, we focus on the overlooked aspect of the noise arising from inhomogeneity of copies of biomolecules in a realistic biological environment. Such inhomogeneity leads to variations of the radical pair parameters, thereby deteriorating the signal arising from an ensemble and providing a source of noise. We investigate the effect of variations in hyperfine interactions between different copies of simple radical pairs on the directional response of a compass system. We find that the choice of radical pair parameters greatly influences how strongly the directional response of an ensemble is affected by inhomogeneity.

Extended Electron-Transfer in Animal Cryptochromes Mediated by a Tetrad of Aromatic Amino Acids

The cryptochrome/photolyase protein family possesses a conserved triad of tryptophans that may act as a molecular wire to transport electrons from the protein surface to the FAD cofactor for activation and/or signaling-state formation. Members from the animal (and animal-like) cryptochrome subclade use this process in a light-induced fashion in a number of exciting responses, such as the (re-)setting of circadian rhythms or magnetoreception; however, electron-transfer pathways have not been explored in detail yet. Therefore, we present an in-depth time-resolved optical and electron-paramagnetic resonance spectroscopic study of two cryptochromes from Chlamydomonas reinhardtii and Drosophila melanogaster. The results do not only reveal the existence of a fourth, more distant aromatic amino acid that serves as a terminal electron donor in both proteins, but also show that a tyrosine is able to fulfill this very role in Chlamydomonas reinhardtii cryptochrome. Additionally, exchange of the respective fourth aromatic amino acid to redox-inactive phenylalanines still leads to light-induced radical pair formation; however, the lifetimes of these species are drastically reduced from the ms- to the μs-range. The results presented in this study open up a new chapter, to our knowledge, in the diversity of electron-transfer pathways in cryptochromes. Moreover, they could explain unique functions of animal cryptochromes, in particular their potential roles in magnetoreception because magnetic-field effects of light-induced radical pairs strongly depend on distance and orientation parameters.

Ready access to end-functional polystyrenes via a combination of ARGET ATRP and thiol–ene chemistry

Bromine end-groups of ATRP polystyrenes were converted into terminal alkene groupsviaa facile synthetic strategy. Subsequently, a light-induced radical thiol–ene reaction was employed for the functionalization of the terminal double bonds.

Cellular metabolites enhance the light sensitivity of Arabidopsis cryptochrome through alternate electron transfer pathways.

Cryptochromes are blue light receptors with multiple signaling roles in plants and animals. Plant cryptochrome (cry1 and cry2) biological activity has been linked to flavin photoreduction via an electron transport chain comprising three evolutionarily conserved tryptophan residues known as the Trp triad. Recently, it has been reported that cry2 Trp triad mutants, which fail to undergo photoreduction in vitro, nonetheless show biological activity in vivo, raising the possibility of alternate signaling pathways. Here, we show that Arabidopsis thaliana cry2 proteins containing Trp triad mutations indeed undergo robust photoreduction in living cultured insect cells. UV/Vis and electron paramagnetic resonance spectroscopy resolves the discrepancy between in vivo and in vitro photochemical activity, as small metabolites, including NADPH, NADH, and ATP, were found to promote cry photoreduction even in mutants lacking the classic Trp triad electron transfer chain. These metabolites facilitate alternate electron transfer pathways and increase light-induced radical pair formation. We conclude that cryptochrome activation is consistent with a mechanism of light-induced electron transfer followed by flavin photoreduction in vivo. We further conclude that in vivo modulation by cellular compounds represents a feature of the cryptochrome signaling mechanism that has important consequences for light responsivity and activation.

A strategy to design biocompatible polymer particles possessing increased loading efficiency and controlled-release properties

Temperature-responsive poly(N-isopropylacrylamide), or poly(NIPAM), layers were reliably prepared around guest molecule (i.e., rhodamine B)-loaded mesoporous silica (SiO2) particles via thermally- and light-induced radical polymerizations. Subsequent removal of the sacrificial SiO2 particles with dilute hydrofluoric acid led to the formation of biocompatible polymer particles possessing a high dose of rhodamine B. The use of SiO2 core templates not only led to the formation of a uniform coating of the poly(NIPAM) layers, but also increased the stability of the guest molecule, rhodamine B, throughout polymerization. Interestingly, the light-induced radical polymerization method resulted in much less inevitable leaching and decomposition of azo-based guest molecules. The structural information and overall dye-loading efficiency of the mesoporous particles and the final polymer particles were then thoroughly examined by electron microscopes, dynamic light scattering, and fluorescence spectroscopy. As poly(NIPAM)-based particles exhibited significant swelling and deswelling properties above and below the lower critical solution temperature, the controlled-release properties of the poly(NIPAM) particles prepared by both methods were also evaluated. Generally, the dye-loaded poly(NIPAM) particles prepared by the light-induced approach resulted in a thinner coating of the polymer layers and exhibited much higher loading and tunable release profiles of the loaded guest molecules than those prepared by the thermally-induced polymerization. Given these features, the generalization of our strategy to design chemotherapeutically interesting drug-loaded polymer particles that are biocompatible and sensitive to external stimuli will allow for the further development of novel biomedical delivery and treatment systems.

One-Dimensional Organic-Inorganic Nanocomposite Synthesized with Single-Walled Carbon Nanotube Templates.

This study reports on single-walled carbon nanotubes (SWCNT) as templates for the preparation of 1D porous organic-inorganic hybrid composites. The in situ deposited SWCNT were sputter coated with Sn metal and thermally oxidized in air to form a SnO2/SWCNT nanowire framework on SiO2/Si substrate. Poly(acrylic acid) (PAA) was coated onto this scaffold through UV light-induced radical polymerization, which resulted in the final formation of hybrid composites. The structures of hybrid composites were investigated by scanning electron microscopy, transmission electron microscopy, infrared spectroscopy, and Raman spectroscopy. The results show that PAA was successfully coated and the structural advantage of nanowire was fairly maintained, which indicates that this framework is very stable for organic functionalization in solution. The simplicity of this method for the formation of porous organic-inorganic hybrid composites provides a potential application for nanoelectronic devices.

Spin relaxation and structure of light-induced spin-correlated PCBM−/P3HT+ radical pairs

The electron spin echo (ESE) technique is applied to determine the spin relaxation times of long-lived lightinduced radicals and short-term spin-correlated radical pairs (SCRPs) formed by the laser flash of a composite consisting of [6,6]-phenyl-С61-butyric acid methyl ether (PCBM) and poly-(3-hexylthiophene) (P3HT) at 80 K. The ESE signal dependences recorded to measure the longitudinal relaxation times of P3HT + /PCBM – SCRPs and the free P3HT + radical are fitted by the exp(–(t/T1) 0.6 ) dependence with Т1 values lying in the microsecond time scale. The difference in the transverse spin relaxation times of the P3HT + /PCBM – radical paira appeared after selective and non-selective echo-detected EPR spectrum excitation is explained by the instantaneous diffusion model. Based on the model, the magnetic interaction energy between the electron spins in P3HT + /PCBM – SCRPs is estimated; E/ћ ∼ 10 6 s –1 .

ATP binding turns plant cryptochrome into an efficient natural photoswitch.

Cryptochromes are flavoproteins that drive diverse developmental light-responses in plants and participate in the circadian clock in animals. Plant cryptochromes have found application as photoswitches in optogenetics. We have studied effects of pH and ATP on the functionally relevant photoreduction of the oxidized FAD cofactor to the semi-reduced FADH· radical in isolated Arabidopsis cryptochrome 1 by transient absorption spectroscopy on nanosecond to millisecond timescales. In the absence of ATP, the yield of light-induced radicals strongly decreased with increasing pH from 6.5 to 8.5. With ATP present, these yields were significantly higher and virtually pH-independent up to pH 9. Analysis of our data in light of the crystallographic structure suggests that ATP-binding shifts the pKa of aspartic acid D396, the putative proton donor to FAD·−, from ~7.4 to >9, and favours a reaction pathway yielding long-lived aspartate D396−. Its negative charge could trigger conformational changes necessary for signal transduction.

Charge Recombination in SnTyrZ•QA–• Radical Pairs in D1 Protein Variants of Photosystem II: Long Range Electron Transfer in the Marcus Inverted Region

Charge recombination in the light-induced radical pair SnTyrZ(•)QA(-•) in Photosystem II (PSII) from Thermosynechococcus elongatus has been studied at cryogenic temperatures by time-resolved EPR for different configurations of PSII that are expected to affect the driving force of the reaction (oxidation states S0, S1, or S2 of the Mn4CaO5 cluster; PsbA1, PsbA2, or PsbA3 as D1 protein). The kinetics were independent of temperature in the studied range from 4.2 to 50 K and were not affected by exchange of H2O for D2O, consistent with single-step electron tunneling over the distance of ∼32 Å without any repopulation through Boltzmann equilibration of intermediates lying higher in energy. In PsbA1-PSII, the charge recombinations in the radical pairs SnTyrZ(•)QA(-•) (ket = 3.4 × 10(-3) s(-1) for S1) were slower than in PsbA3-PSII despite an expected lower driving force owing to a downshifted Em(QA/QA(-•)) in PsbA1-PSII. Conversely, the reaction was slower in the presence of S2 than in the presence of S1, despite an expected larger driving force due to an upshifted Em(TyrZ(•)/TyrZ) in S2. These observations indicate that the charge recombination occurs in the Marcus inverted region. Assuming that the driving force of the reaction (-ΔG(0) ≈ 1.2 eV at room temperature for S1) does not vary strongly with temperature, the data indicate an optimal electron transfer rate (for a hypothetical -ΔG(0) = λ) substantially faster than would be predicted from extrapolation of room temperature intraprotein ET rates over shorter distances. Possible origins of this deviation are discussed, including a possible enhancement of the electronic coupling of TyrZ(•) and QA(-•) by aromatic cofactors located in between. Observed similar S1TyrZ(•)QA(-•) charge recombinations in PsbA2-PSII and PsbA3-PSII predict that Em(QA/QA(-•)) in PsbA2-PSII is similar to that in PsbA3-PSII.

Highly Branched Polyisobutylene by Radical Polymerization under Li[CB11(CH3)12] Catalysis

In the presence of a nonoxidizing radical initiator, azo-tert-butane, and a high concentration of LiCB11(CH3)12, isobutylene undergoes thermal or light-induced radical polymerization to b-PIB, a hi...

Light-induced Free Radical Oxidation of 2-Oxo-1,2,3,4-tetrahydropyrimidines

Light-induced Free Radical Oxidation of 2-Oxo-1,2,3,4-tetrahydropyrimidines

Hierarchically designed injectable hydrogel from oxidized dextran, amino gelatin and 4-arm poly(ethylene glycol)-acrylate for tissue engineering application

Hydrogels are high in water content and have physical properties similar to native extracellular matrix (ECM), and thus they have been widely studied as three-dimensional (3D) tissue engineering scaffolds for cell culture. In this work, a two-step process was introduced to fabricate injectable hydrogel from oxidized dextran (ODex), amino gelatin (MGel) and 4-arm poly(ethylene glycol)-acrylate (4A-PEG-Acr) for cell encapsulation. A primary network was formed based on a Schiff based reaction between ODex and MGel, then a UV light-induced radical reaction of 4A-PEG-Acr was used to produce the independent secondary network. Both of the reactions were carried out under physiological conditions in the presence of living cells with no toxicity. The primary network depending on natural polymers could degrade rapidly to provide space and nutrition for encapsulated cells’ growth, and the secondary network could provide long-term mechanical stability. The attachment and spreading of pre-osteoblasts (MC3T3-E1) on IPN hydrogels were observed by DEAD/LIVE kit staining. Furthermore, cell spreading and cell proliferation within IPN hydrogels were observed using confocal microscopy after phalloidin/DAPI staining. The results showed that the as-prepared interpenetrating polymer network (IPN) hydrogels possessed good mechanical properties, a controllable degradation rate and favorable biocompatibility. Therefore, the hierarchically designed hydrogel in this study could be a promising candidate for bone or cartilage tissue engineering applications.

Neutral metallated and meso-substituted porphyrins as antimicrobial agents against Gram-positive pathogens

Staphylococcus aureus is a bacterial pathogen that causes severe infections among humans. The increasing emergence of antibiotic resistance necessitates the development of new strategies to combat the spread of disease. One approach is photodynamic inactivation using porphyrin photosensitizers, which generate superoxide and other radicals in the presence of light, causing cell death via the oxidation of proteins and lipids. In this study, we analyzed a novel library of meso-substituted and metallated porphyrins for activity against multidrug-resistant S. aureus. From a library of 251 compounds, 51 showed antimicrobial activity, in three discrete classes of activity: those that functioned only in light, those that had toxicity only in darkness, and those that displayed activity regardless of illumination. We further demonstrated the broad-spectrum activity of these compounds against a variety of pathogens, including Bacillus anthracis, Enterococcus faecalis, and Escherichia coli. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) analyses of lead compounds (XPZ-263 and XPZ-271) revealed strong activity and killing towards methicillin-resistant S. aureus (MRSA) strains. An analysis of mutation frequencies revealed low incidences of resistance to lead compounds by E. coli and MRSA. Finally, an exploration of the underlying mechanism of action suggests that these compounds do not depend solely upon light-induced radical generation for toxicity, highlighting their potential for clinical applications.

Quantum effects in biology: Bird navigation

Birds possess a physiological magnetic compass, enabling them to make correct directional choices during their migratory flights. The biophysical basis of this compass remains unknown. One of the two major hypotheses postulates that birds use a light-induced radical pair reaction involving coherent spin evolution of two electrons as the foundation of their magnetic compass sensor. This hypothesis has received significant attention during the past decade. Here, we review the growing body of chemical, biophysical, behavioral, and neurobiological evidence suggesting that this quantum-based mechanism may indeed form the basis of the avian magnetic compass.

3P278 シロイヌナズナFKF1タンパクLOVドメインにおいて低温光反応で検出されるアニオンラジカルのEPR測定(光生物-視覚・光受容,第48回日本生物物理学会年会)

3P278 シロイヌナズナFKF1タンパクLOVドメインにおいて低温光反応で検出されるアニオンラジカルのEPR測定(光生物-視覚・光受容,第48回日本生物物理学会年会)

Photocatalytic Generation of Oxygen Radicals by the Water-Soluble Bacteriochlorophyll Derivative WST11, Noncovalently Bound to Serum Albumin

Light-induced radical generation is the hallmark of fundamental processes and many applications including photosynthesis and photodynamic therapy (PDT). In this manuscript, we present two novel observations made upon monitoring light-induced generation of reactive oxygen species (ROS) in aqueous solutions by WST11, a water-soluble derivative of the photosynthetic pigment Bacteriochlorophyll a (Bchl). Using a host of complementary experimental techniques including time-resolved spectroscopy at the subpicosecond to the millisecond range, ESR spectroscopy, electrochemistry, spectroelectrochemistry, oximetry, and protein mass spectroscopy, we first show that in aqueous solutions WST11 generates only superoxide (O(2)(-*)) and hydroxyl (OH*) radicals with no detectable traces of singlet oxygen. Second, we show that WST11 makes a noncovalent complex with human serum albumin (HSA) and that this complex functions as a photocatalytic oxidoreductase at biologically relevant concentrations enabling approximately 15 cycles of electron transfer from the associated HSA protein to molecular oxygen in the solution. These findings rule out the paradigm that porphyrin and chlorophyll based PDT is mainly mediated by formation of singlet oxygen, particularly in vascular targeted photodynamic therapy (VTP) with sensitizers that undergo photoactivation during circulation in the plasma, like [Pd]-Bacteriopheophorbide (WST09, Tookad). At the same time, our findings open the way for new design paradigms of novel sensitizers, since O(2)(-*) and OH* radicals are well-recognized precursors of important pathophysiological processes that can be activated for achieving tumor eradication. Moreover, the finding that promiscuous protein scaffolds become sinks for holes and electrons when holding light-activated pigments provides a new insight to the evolution and action mechanism of natural light activated oxidoreductases (such as photosynthetic reaction centers) and new guidelines for the preparation of synthetic-light converting machineries.

What you get out of high-time resolution electron paramagnetic resonance: example from photosynthetic bacteria

The primary energy conversion steps of natural photosynthesis proceed via light-induced radical ion pairs as short-lived intermediates. Time-resolved electron paramagnetic resonance (EPR) experiments of photosynthetic reaction centers monitor the key charge separated state between the oxidized primary electron donor and reduced quinone acceptor, e.g., P(+)(865)Q(-)(A) of purple photosynthetic bacteria. The time-resolved EPR spectra of P(+)(865)Q(-)(A) are indicative of a spin-correlated radical pair that is created from the excited singlet state of P(865) in an ultra-fast photochemical reaction. Importantly, the spin-correlated radical pair nature of the charge separated state is a common feature of all photosynthetic reaction centers, which gives rise to several interesting spin phenomena such as quantum oscillations, observed at short delay times after optical excitation. In this review, we describe details of the quantum oscillation phenomenon and present a complete analysis of the data obtained from the charge separated state of purple bacteria, P(+)(865)Q(-)(A). The analysis and simulation of the quantum oscillations yield the three-dimensional structure of P(+)(865)Q(-)(A) in the photosynthetic membrane on a nanosecond time scale after light-induced charge separation. Comparison with crystallographic data reveals that the position of Q(-)(A) is essentially the same as in the X-ray structure. However, the head group of Q(-)(A) has undergone a 60° rotation in the ring plane relative to its orientation in the crystal structure. The results are discussed within the framework of the previously suggested conformational gating mechanism for electron transfer from Q(-)(A) to the secondary quinone acceptor Q(B).