Protonation State Research Articles

Unusual variability of isomers in copper(II) complexes with 1,8-bis(2-hydroxybenzyl)-cyclam.

Copper isotopes and their complexes are intensively studied due to their high potential for applications in radiodiagnosis and radiotherapy. Here, we study the CuII complex of 1,8-bis(2-hydroxybenzyl)-cyclam (H2L), which forms an unexpected variety of isomers differing in the mutual orientation of the substituents on the cyclam nitrogen atoms, the protonation of the phenolate pendant, and the ligand denticity. The interconversion of the isomers is rather slow, which made the isolation, identification and investigation of some of the individual species possible. The most stable and the most common form is the hexacoordinated trans-III isomer. However, several other forms were also observed in solution in the course of HPLC, UV-VIS and electrochemical measurements. The isomers present in solution were identified by comparison with the solid-state structures solved by X-ray diffraction analysis on single crystals and with the help of theoretical calculations. The phenolate pendant is coordinated both in the protonated and deprotonated state; however, the coordination in the axial position of the hexacoordinated trans-III complex is weak, especially in its protonated state. Conversely, the CuII ion is pentacoordinated in the cis-V isomer with only one phenolate strongly coordinated in the basal plane of the distorted tetragonal pyramid. The computational data showed that the phenolate groups might form strong intraligand hydrogen bonds competitive with the metal-phenolate bonds, stabilizing the structure of the complex. In addition, theoretical calculations revealed that several geometries are energetically close to the optimal one, which indicates possible dynamic behaviour of the complex in solution.

Measurement of tt¯ production in association with additional b-jets in the eμ final state in proton–proton collisions at s = 13 TeV with the ATLAS detector

This paper presents measurements of top-antitop quark pair (tt¯) production in association with additional b-jets. The analysis utilises 140 fb−1 of proton–proton collision data collected with the ATLAS detector at the Large Hadron Collider at a centre-of-mass energy of 13 TeV. Fiducial cross-sections are extracted in a final state featuring one electron and one muon, with at least three or four b-jets. Results are presented at the particle level for both integrated cross-sections and normalised differential cross-sections, as functions of global event properties, jet kinematics, and b-jet pair properties. Observable quantities characterising b-jets originating from the top quark decay and additional b-jets are also measured at the particle level, after correcting for detector effects. The measured integrated fiducial cross-sections are consistent with tt¯bb¯ predictions from various next-to-leading-order matrix element calculations matched to a parton shower within the uncertainties of the predictions. State-of-the-art theoretical predictions are compared with the differential measurements; none of them simultaneously describes all observables. Differences between any two predictions are smaller than the measurement uncertainties for most observables.

Protonation effects in protein-ligand complexes - a case study of endothiapepsin and pepstatin A with computational and experimental methods.

Protonation states serve as an essential molecular recognition motif for biological processes. Their correct consideration is key to successful drug design campaigns, since chemoinformatic tools usually deal with default protonation states of ligands and proteins and miss atypical protonation states. The protonation pattern for the Endothiapepsin/PepstatinA (EP/pepA) complex is investigated using different dry lab and wet lab techniques. ITC experiments revealed an uptake of more than one mole of protons upon pepA binding to EP. Since these experiments were performed at physiological conditions (and not at pH=4 at which a large variety of crystal structures is available), a novel crystal structure at pH=7.6 was determined. This crystal structure showed that only modest structural changes occur upon increasing the pH value. This lead to computational studies (Poisson Boltzmann calculations and constant pH MD simulations)to reveal the exact location of the protonation event. Both computational studies could reveal a significant pKa shift resulting in non-default protonation state and that the catalytic dyad is responsible for the uptake of protons. This study shows that assessing protonation states for two separate systems (protein and ligand) might result in the incorrect assignment of protonation states and hence incorrect calculation of binding energy.

Computational Chemistry Study of pH-Responsive Fluorescent Probes and Development of Supporting Software

This study employs quantum chemical computational methods to predict the spectroscopic properties of fluorescent probes 2,6-bis(2-benzimidazolyl)pyridine (BBP) and (E)-3-(2-(1H-benzo[d]imidazol-2-yl)vinyl)-9-(2-(2-methoxyethoxy)ethyl)-9H-carbazole (BIMC). Using time-dependent density functional theory (TDDFT), we successfully predicted the fluorescence emission wavelengths of BBP under various protonation states, achieving an average deviation of 6.0% from experimental excitation energies. Molecular dynamics simulations elucidated the microscopic mechanism underlying BBP’s fluorescence quenching under acidic conditions. The spectroscopic predictions for BIMC were performed using the STEOM-DLPNO-CCSD method, yielding an average deviation of merely 0.57% from experimental values. Based on Einstein’s spontaneous emission formula and empirical internal conversion rate formulas, we calculated fluorescence quantum yields for spectral intensity calibration, enabling the accurate prediction of experimental spectra. To streamline the computational workflow, we developed and open-sourced the EasySpecCalc software v0.0.1 on GitHub, aiming to facilitate the design and development of fluorescent probes.

Ring-in-Ring Assembly Facilitates the Synthesis of a [12]Cycloparaphenylene ABC-Type [3]Catenane.

Cycloparaphenylenes (CPPs) represent a significant challenge for the synthesis of mechanically interlocked architectures, because they lack heteroatoms, which precludes traditional active and passive template methods. To circumvent this problem and explore the fundamental and functional properties of CPP rotaxanes and catenanes, researches have resorted to unusual non-covalent and even to labor-intensive covalent template approaches. Herein, we report a ring-in-ring non-covalent template strategy that makes use of the surprisingly strong non-covalent inclusion of crown ethers into suitably sized CPPs. By threading a secondary ammonium salt through the crown ether and closing the third ring via CuAAC click reaction, we obtained a rare ABC-type hetero-[3]catenane comprising [12]CPP, 24-crown-8 and a dibenzylammonium macrocycle. X-ray crystallography shed light on the ring-in-ring pre-organization and the [3]catenane topology was confirmed by NMR and MS-MS studies. Molecular simulations provided insights into the intriguing ring-vs.-ring-vs.-ring dynamics of the [3]catenane, which are highly dependent on the protonation state of the dibenzylammonium site. This ring-in-ring assembly strategy opens new avenues for the synthesis of complex CPP architectures and their use in functional supramolecular systems.

Additive CHARMM Force Field for Pterins and Folates.

Folates comprise a crucial class of biologically active compounds related to folic acid, playing a vital role in numerous enzymatic reactions. One-carbon metabolism, facilitated by the folate cofactor, supports numerous physiological processes, including biosynthesis, amino acid homeostasis, epigenetic maintenance, and redox defense. Folates share a common pterin heterocyclic ring structure capable of undergoing redox reactions and existing in various protonation states. This study aimed to derive molecular mechanics (MM) parameters compatible with the CHARMM36 all-atom additive force field for pterins and biologically important folates, including pterin, biopterin, and folic acid. Three redox forms were considered: oxidized, dihydrofolate, and tetrahydrofolate states. Across all protonation states, a total of 18 folates were parameterized. Partial charges were derived using the CHARMM force field parametrization protocol, based on targeting reference quantum mechanics monohydrate interactions, electrostatic potential, and dipole moment. Bonded terms were parameterized using one-dimensional adiabatic potential energy surface scans, and two-dimensional scans to parametrize in-ring torsions associated with the puckering states of dihydropterin and tetrahydropterin. The quality of the model was demonstrated through simulations of three protein complexes using optimized and initial parameters. These simulations underscored the significantly enhanced performance of the folate model developed in this study compared to the initial model without optimization in reproducing structural properties of folate-protein complexes. Overall, the presented MM model will be valuable for modeling folates in various redox states and serve as a starting point for parameterizing other folate derivatives.

Identification and characterization of shifted G•U wobble pairs resulting from alternative protonation of RNA.

RNA can serve as an enzyme, small molecule sensor, and vaccine, and it may have been a conduit for the origin of life. Despite these profound functions, RNA is thought to have quite limited molecular diversity. A pressing question, therefore, is whether RNA can adopt novel molecular states that enhance its function. Covalent modifications of RNA have been demonstrated to augment biological function, but much less is known about non-covalent alterations such as novel protonated or tautomeric forms. Conventionally, a G•U wobble has the U shifted into the major groove. We used a cheminformatic approach to identify four structural families of shifted G•U wobbles in which the G instead resides in the major groove of RNA, which requires alternative tautomeric states of either base, or an anionic state of the U. We provide experimental support for these shifted G•U wobbles via the potent, and unconventional, in vivo reactivity of the U with dimethylsulfate (DMS) in three organisms. These shifted wobbles may play important functional roles and could serve as drug targets. Our cheminformatics approach is general and can be applied to identify alternative protonation states in other RNA motifs, as well as in DNA and proteins.

An albumin unfolding and refolding cycle induced by a time-controlled pH jump.

Given the intimate connection between the structure and function of biological macromolecules, the ability to temporally control their unfolding-refolding process enables temporal regulation over specific functionalities, potentially applicable in innovative domains, including the construction of protein-based actuators or programmable catalysis and drug release in complex biotechnological processes. We show here how a temporally controlled protein unfolding-refolding cycle can be coupled in time with programmed pH sequences achieved through the spontaneous decomposition of an activated carboxylic acid. Specifically, we illustrate this process at the molecular level using albumin, the most prevalent protein found in plasma, for which a temporary shift from native to unfolded forms is promoted using nitroacetic acid, able to undergo base-catalysed decarboxylation when solubilized in water solution. As detected by small angle X-ray scattering and intrinsic tryptophan fluorescence, starting from the protein in its native form, the acid addition triggers unfolding to a partially denatured state and a subsequent time-tunable pH rise with gradual refolding that recapitulates the intermediate steps detected at the same pH values by static acidification, fitting within a framework of full reversibility of the structural changes as a function of the protein protonation state. At the end of the pH jump, the native structure is fully recovered, making this method a chemical tool to achieve a complete protein conformational sequence programmed in the timeframe of minutes without further intervention.

Nanofibrous Peptide Hydrogels Leveraging Histidine to Modulate pH-Responsive Supramolecular Assembly and Antibody Release.

In this work, we investigate the pH-responsive behavior of multidomain peptide (MDP) hydrogels containing histidine. Small-angle X-ray scattering confirmed that MDP nanofibers sequester nonpolar residues into a hydrophobic core surrounded by a shell of hydrophilic residues. MDPs with histidine on the hydrophilic face formed nanofibers at all pH values tested, but the morphology of the fibers was influenced by the protonation state and the location of histidine in the MDP sequence. MDPs with histidine residues within the hydrophobic face disassemble below physiological pH and form nanofibers at higher pH. Taking advantage of their stimulus-triggered behavior, an anti-PD-1 antibody was loaded into histidine MDP hydrogels to examine pH-dependent differences in payload delivery in vitro. Hydrogels composed of MDPs with histidine on the hydrophilic face demonstrated pH-dependent payload retention. Additionally, they showed significantly slower antibody release and reduced antibody diffusion rates in vitro compared to MDP hydrogels lacking histidine.

Proton Occupancies in Histidine Side Chains of Carbonic Anhydrase II by Neutron Crystallography and NMR - Differences, Similarities and Opportunities.

Histidine is a key amino-acid residues in proteins that can exist in three different protonation states: two different neutral tautomeric forms and a protonated, positively charged one. It can act as both donor and acceptor of hydrogen bonds, coordinate metal ions, and engage in acid/base catalysis. Human Carbonic Anhydrase II (HCA II) is a pivotal enzyme catalyzing the reversible hydration of carbon dioxide. It contains 12 histidine residues: sixsurface exposed, two buried, three active site zinc ion ligands, and one is a proton shuttle. Comparing results from NMR spectroscopy with previously determined neutron protein crystal structures enabled a side-by-side investigation of the proton occupancies and preferred tautomeric states of the histidine residues in HCA II. Buried and zinc coordinating histidines remain in one neutral tautomeric state across the entire pH range studied, as validated by both methods. In contrast, solvent-exposed histidines display high variability in proton occupancies. While the data were overall remarkably consistent between methods, some discrepancies were observed, shedding light on the limitations of each technique. Therefore, combining these methods with full awareness of the advantages and drawbacks of each, provides insights into the dynamic protonation landscape of HCA II histidines, crucial for elucidating enzyme catalytic mechanisms.

Structural Insights Into the Opening Mechanism of C1C2 Channelrhodopsin.

Channelrhodopsins, light-gated cation channels, enable precise control of neural cell depolarization or hyperpolarization with light in the field of optogenetics. This study integrates time-resolved serial crystallography and atomistic molecular dynamics (MD) simulations to resolve the structural changes during C1C2 channelrhodopsin activation. Our observations reveal that within the crystal environment, C1C2 predominantly remains in a light-activated state with characteristics of the M390 intermediate. Here, rearrangement of retinal within its binding pocket partially opens the central gate toward the extracellular vestibule. These structural changes initiate channel opening but were insufficient to allow K+ flow. Adjusting protonation states to represent the subsequent N520 intermediate in our MD simulations induced further conformational changes, including rearrangements of transmembrane helices 2 and 7, that opened the inner gate and the putative ion-translocation pathway. This allowed spontaneous cation conduction with low conductance, aligning with experimental findings. Our findings provide critical structural insights into key intermediates of the channel opening mechanism, enhancing our understanding of ion conduction and selectivity in channelrhodopsins at an atomistic level.

Organic Crystals: Comprehensive Understanding of Proton States of Naftopidil Salts via XPS

Organic Crystals: Comprehensive Understanding of Proton States of Naftopidil Salts via XPS

Biliverdin's Propionic Chains Influence Oligomerization in Sandercyanin.

Sandercyanin is a mildly fluorescent biliprotein with a large Stokes shift, a tetrameric quaternary structure, and a biliverdin (BV) chromophore that does not covalently bond to the protein. To adapt this promising protein for use in bioimaging, it is necessary to produce monomeric mutants that retain the spectroscopic properties while increasing the fluorescence quantum yield. Modulating these properties through the protonation state of BV's propionic tails is a possible avenue, if detailed mechanistic information on the role of such chains becomes available. In this study, we use a microstate model for the titration process of BV and couple it with constant pH molecular dynamics to study protonation states in the apo protein, the artificial monomer, and the tetramer and identify shifts. Our results indicate that several residues might have a central role in oligomerization as a response to the presence of BV and especially to the protonation state of the propionic tails. While the absorption properties are not strongly impacted by the tails, their protonation state has an impact on the chromophore geometry, which likely influences the fluorescence.

Low-frequency Raman spectroscopy as a new tool for understanding the behaviour of ionisable compounds in dispersed mesophases.

Low-frequency Raman (LFR) spectroscopy is proposed as a novel non-destructive methodology to probe pH-related phase transitions in self-assembled lipid particles. In this case, dispersed lipid mesophases were composed of ionisable oleic acid (OA) or nicergoline (NG) in monoolein (MO). The sensitivity of LFR spectroscopy to low-energy intermolecular vibrations was hypothesised to be due to structural transformation in ionisable dispersed mesophases upon changes in pH. Phase transitions of dispersed mesophases of MO mixed with OA or NG were induced by varying the pH of the aqueous buffer. The structural transformations were studied using LFR spectroscopy, recording the corresponding changes in the vibrational density of states (VDOS) upon changes in pH and analysed using principal component analysis (PCA). The results were correlated with structural transitions observed in simultaneous small-angle X-ray scattering (SAXS) measurements. The intensity of the VDOS signal of MO+OA mesophases scaled with phase-specific transformations, such as from the bi-continuous cubic Im3¯m phase (V2) or lamellar-based vesicles to the reversed hexagonal p6m phase (H2). For NG subtle changes in the lattice parameter of the V2 phase of NG+MO mesophases coincided with the apparent dissociation constant (pKaapp) of NG, however, slight variations between the pKaapp of NG determined by equilibrated samples analysed using SAXS and non-equilibrated samples analysed using LFR suggest structural hysteresis upon changes in the protonation state of NG. This approach offers an efficient method for studying the phase behaviour of lipid systems under varying pH and potentially other conditions such as temperature.

Antimicrobial Activity of Glycyrrhizinic Acid Is pH-Dependent.

In recent years, antimicrobial hydrogels have attracted much attention in biomedical applications due to their biocompatibility and high water content. Glycyrrhizin (GA) is an antimicrobial that can form pH-dependent hydrogels due to the three carboxyl groups of GA that differ in pKa value. The influence of GA protonation on the antimicrobial activity, however, has never been studied before. Therefore, we investigated the effect of the pH on the antimicrobial activity of GA against Pseudomonas aeruginosa, Staphylococcus aureus, MRSA, Staphylococcus epidermidis, Acinetobacter baumannii, Klebsiella pneumoniae, Klebsiella aerogenes, and two strains of Escherichia coli. In general, the antimicrobial activity of GA increases as a function of decreasing pH (and thus increasing protonation of GA). More specifically, fully protonated GA hydrogels (pH = 3) are required for growth inhibition and killing of E. coli UTI89 and Klebsiella in the suspension above the hydrogel, while the staphylococci strains and A. baumannii are already inhibited by fully deprotonated GA (pH = 6.8). P. aeruginosa and E. coli DH5α showed moderate susceptibility, as they are completely inhibited by a hydrogel at pH 3.8, containing partly protonated GA, but not by fully deprotonated GA (pH = 6.8). The antimicrobial activity of the hydrogel cannot solely be attributed to the resulting pH decrease of the suspension, as the presence of GA significantly increases the activity. Instead, this increased activity is due to the release of GA from the hydrogel into the suspension, where it directly interacts with the bacteria. Moreover, we provide evidence indicating that the pH dependency of the antimicrobial activity is due to differences in GA protonation state by treating the pathogens with GA solutions differing in their GA protonation distribution. Finally, we show by LC-MS that there is no chemical or enzymatic breakdown of GA. Overall, our results demonstrate that the pH influences not only the physical but also the antimicrobial properties of the GA hydrogels.

Charge-Selective Electrochemical Detection through Electrolyte Adsorption on Indium Tin Oxide Electrodes

In the field of electroanalytical methods, selective sensing of target analytes is as crucial as sensitive detections, as interfering substances coexist in real samples. Therefore, various strategies have been developed to enhance the selectivity of electrochemical sensors, including molecularly imprinted polymers, enzymes, or nanostructured electrodes. Modifying the charge of electrode have been also developed, which leads to charge-based discrimination, and eventually resulting in selective electrochemical sensing of charged species. For example, the introduction of negatively charged functional groups (e.g., carboxyl groups) on electrodes has shown selective detection towards positively charged species (e.g., dopamine). However, such approaches require complex materials that are customized for specific target redox species, which limits their wide applicability. In this work, we utilized the adsorption of electrolyte anions on bare indium tin oxide (ITO) electrodes to control the surface charge of the electrode and thus enable charge-based discrimination of redox species. By using polyprotic acid anions as electrolytes and varying the solution pH to determine their protonation states, we could systematically assess the electrochemical response of adsorbed charge density towards negative, positive, and neutral redox species. Furthermore, the anion effect was applied to the selective detection of dopamine in the presence of the two representative interfering species in blood, ascorbic acid and uric acid. We expect that the electrolyte-derived surface charging of the ITO electrode could be a widely applicable strategy for the selective detection of charged analytes.

Selective Protonation of Catalytic Dyad for γ-Secretase-Mediated Hydrolysis Revealed by Multiscale Simulations.

γ-Secretase plays a crucial role in producing disease-related amyloid-β proteins by cleaving the amyloid precursor protein (APP). The enzyme employs its catalytic dyad containing two aspartates (Asp257 and Asp385) to hydrolyze the substrate by a general acid-base catalytic mechanism, necessitating monoprotonation of the two aspartates for efficient hydrolysis. However, the precise protonation states of the aspartates remain uncertain. In this study, we employed a multiscale computational approach to investigate the dependence of the catalytic efficiency of γ-secretase on the protonation states of its catalytic dyad. Over 200 ms unbiased atomistic simulations of the substrate-enzyme complex reveal diverse orientations of the scissile bond of the bound substrate and accessible structural ensembles of the catalytic dyad with Asp257-Asp385 distances fluctuating between 4 and 10 Å. With a quantum mechanics/molecular mechanics (QM/MM) approach accelerated by enhanced sampling techniques, we find that the first step of the hydrolysis reaction, i.e., the formation of a gem-diol intermediate, experiences a higher reaction barrier by ∼2 kcal/mol when Asp385 is protonated. Furthermore, we find that Arg269 of the enzyme is most likely responsible for this preference of the protonation state: its basic side chain is spatially close to that of Asp257 and specifically stabilizes the transition state electrostatically when Asp257 is protonated. Collectively, our study suggests that Asp257 is likely the favored protonation site for APP cleavage by γ-secretase.

Data-Efficient Active Learning for Thermodynamic Integration: Acidity Constants of BiVO4 in Water.

The protonation state of molecules and surfaces is pivotal in various disciplines, including (electro-)catalysis, geochemistry, biochemistry, and pharmaceutics. Accurately and efficiently determining acidity constants is critical yet challenging, particularly when explicitly considering the electronic structure, thermal fluctuations, anharmonic vibrations, and solvation effects. In this research, we employ thermodynamic integration accelerated by committee Neural Network potentials, training a single machine learning model that accurately describes the relevant protonated, deprotonated, and intermediate states. We investigate two deprotonation reactions at the BiVO4 (010)-water interface, a promising candidate for efficient photocatalytic water splitting. Our results illustrate the convergence of the required ensemble averages over simulation time and of the final acidity constant as a function of the Kirkwood coupling parameter. We demonstrate that simulation times on the order of nanoseconds are required for statistical convergence. This time scale is currently unachievable with explicit ab-initio molecular dynamics simulations at the hybrid DFT level of theory. In contrast, our machine learning workflow only requires a few hundred DFT single point calculations for training and testing. Exploiting the extended time scales accessible, we furthermore asses the effect of commonly applied bias potentials. Thus, our study significantly advances calculating free energy differences with ab-initio accuracy.

Atherosclerotic Oxidized Lipids Affect Formation and Biophysical Properties of Supported Lipid Bilayers and Simulated Membranes.

Oxidized lipids arising from oxidative stress are associated with many serious health conditions, including cardiovascular diseases. For example, KDdiA-PC and KOdiA-PC are two oxidized phosphatidylcholines (oxPC) directly linked to atherosclerosis, which precipitate heart failure, stroke, aneurysms, and chronic kidney disease. These oxPCs are well-characterized in small particles such as low-density lipoprotein, but how their presence affects the biophysical properties of larger bilayer membranes is unclear. It is also unclear how membrane mediators, such as cholesterol, affect lipid bilayers containing these oxPCs. Here, we characterize supported lipid bilayers (SLBs) containing POPC, KDdiA-PC, or KOdiA-PC, and cholesterol. We used a quartz crystal microbalance with dissipation monitoring (QCM-D), fluorescence microscopy, and all-atom molecular dynamics (MD) to examine the formation process, biophysical properties, and specific lipid conformations in simulated bilayers. Experimentally, we show that liposomes containing either oxPC form SLBs by rupturing on contact with SiO2 substrates, which differs from the typical adsorption-rupture pathway observed with nonoxidized liposomes. We also show that increasing the oxPC concentration in SLBs results in thinner bilayers that contain defects. Simulations reveal that the oxidized sn-2 tails of KDdiA-PC and KOdiA-PC bend out of the hydrophobic membrane core into the hydrophilic headgroup region and beyond. The altered conformations of these oxPC, which are affected by cholesterol content and protonation state of the oxidized functional groups, contribute to trends of decreasing membrane thickness and increasing membrane area with increasing oxPC concentration. This combined approach provides a comprehensive view of the biophysical properties of membranes containing KDdiA-PC and KOdiA-PC at the molecular level, which is crucial to understanding the role of lipid oxidation in cardiovascular disease and related immune responses.

Computational Characterization of the DAD Photoisomerization: Functionalization, Protonation, and Solvation Effects.

Photoswitches are becoming increasingly popular in pharmacology due to the possibility of modifying their activity with light. Hence, it is crucial to understand the photophysics of these compounds to identify promising light-activated drugs. We focused our study on DAD, an azobenzene derivative that, according to a previous experimental investigation, can restore visual function in blind mice due to trans-cis photoisomerization upon light absorption. With the present computational study, we aim to characterize the absorption spectrum of DAD, and to understand its photoisomerization mechanism by means of conformational search analysis, quantum mechanical (QM) and hybrid QM/continuum calculations, and classical molecular dynamics simulations. Moreover, we explored the effect of the derivation (DAD vs azobenzene), the protonation (DAD vs DADH22+, the two possible protonation states) and the solvation (vacuum vs water) on the photoisomerization. Similarly to azobenzene, we showed that the photoisomerization of both protonation states of DAD begin with the population of the bright S2 state. Then, it crosses to the S1 surface and relaxes along the rotation of the azo dihedral to a S1/S0 crossing point. The latter is close to a transition state that connects the trans and cis geometries on the ground state. Finally, our results suggested that amino derivation, nonprotonation and water solvation could improve the quantum yield of the photoisomerization.