RESUMEN
Expression of concern for 'Direct observation of the THz Kerr effect (TKE) in deionized, distilled and buffered (PBS) water' by Andrzej Dobek et al., Phys. Chem. Chem. Phys., 2017, 19, 26749-26757, https://doi.org/10.1039/c7cp04061j.
RESUMEN
Nonlinear THz pump-optical probe (THz-OKE) measurements in deionized, distilled and buffered (PBS) water are reported. A laser system that produces pulses at 800 nm, 30 fs FWHM, at a repetition rate of 1 kHz and an energy of 6 µJ per pulse, was a source of the probe optical beam and of the pump beam of THz 2 ps pulses. The electric field strength inducing birefringence in water samples was ETHz = 1.35 × 106 V m-1. These samples were chosen in order to study the effect of ionic concentration on water behavior in the ultrafast time scale. Differentiation between ultrafast effects resulting from internal H2O properties and those resulting from H2O-ion interactions was analyzed. These two effects may be connected to a difference in the fluctuations of the network of intermolecular hydrogen bonds of water molecules in the presence and absence of ions in solution. The results indicate that such fluctuations significantly alter water birefringence amplitude and its dynamics.
RESUMEN
The temperature dependence of charge recombination of the pair P(+)HA(-) in isolated reaction centers from the purple bacterium Rhodobacter sphaeroides with prereduced quinone QA was studied by sub-nanosecond to microsecond time-scale transient absorption. Overall, the kinetics slowed down substantially upon cooling from room temperature to â¼200 K, and then remained virtually unchanged down to 77 K, indicating the coexistence of two competitive pathways of charge recombination, a thermally-activated pathway appearing only above ~200 K and a temperature-independent pathway. In our modelling, the thermally activated pathway includes an uphill electron transfer from HA(-) to BA(-) leading to transient formation of the state P(+)BA(-), whereas the temperature-independent pathway is due to direct downhill electron transfer from HA(-) to P(+). At all temperatures studied, the kinetics could be approximated by a four-component decay. Detailed analysis of the lifetimes and amplitudes of particular phases over the range of temperatures suggests that the kinetically resolved phases reveal the consecutive appearance of three conformational states characterized by an increasing free energy gap between the states P(+)BA(-) and P(+)HA(-). The initial gap between these states was estimated to be only ~8 meV, the intermediate gap being ~92 meV, and the final gap ~135 meV, with no dependence on temperature. It was also calculated through a very straightforward approach that the relaxation process from the initial to the intermediate state occurs within 0.6 ± 0.1 ns, whereas the second step of relaxation from the intermediate to the final state takes 11 ± 2 ns. Both phases of the protein relaxation process are essentially temperature-independent. Possible alternative models to describe the experimental data that cannot be definitely excluded are also discussed.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Benzoquinonas/metabolismo , Cinética , Modelos Moleculares , Fenantrolinas/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Temperatura , Termodinámica , Factores de TiempoRESUMEN
Time-resolved spectroscopic studies of recombination of the P(+)HA(-) radical pair in photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides give an opportunity to study protein dynamics triggered by light and occurring over the lifetime of P(+)HA(-). The state P(+)HA(-) is formed after the ultrafast light-induced electron transfer from the primary donor pair of bacteriochlorophylls (P) to the acceptor bacteriopheophytin (HA). In order to increase the lifetime of this state, and thus increase the temporal window for the examination of protein dynamics, it is possible to block forward electron transfer from HA(-) to the secondary electron acceptor QA. In this contribution, the dynamics of P(+)HA(-) recombination were compared at a range of temperatures from 77 K to room temperature, electron transfer from HA(-) to QA being blocked either by prereduction of QA or by genetic removal of QA. The observed P(+)HA(-) charge recombination was significantly slower in the QA-deficient RCs, and in both types of complexes, lowering the temperature from RT to 77 K led to a slowing of charge recombination. The effects are explained in the frame of a model in which charge recombination occurs via competing pathways, one of which is thermally activated and includes transient formation of a higher-energy state, P(+)BA(-). An internal electrostatic field supplied by the negative charge on QA increases the free energy levels of the state P(+)HA(-), thus decreasing its energetic distance to the state P(+)BA(-). In addition, the dielectric response of the protein environment to the appearance of the state P(+)HA(-) is accelerated from â¼50-100 ns in the QA-deficient mutant RCs to â¼1-16 ns in WT RCs with a negatively charged QA(-). In both cases, the temperature dependence of the protein dynamics is weak.
Asunto(s)
Proteínas Bacterianas/química , Bacterioclorofilas/química , Feofitinas/química , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Rhodobacter sphaeroides/metabolismo , Ubiquinona/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Transporte de Electrón , Mutación , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Espectrometría de Fluorescencia , Electricidad Estática , Temperatura , Factores de Tiempo , Ubiquinona/metabolismoRESUMEN
The kinetics of recombination of the P(+)H(A)(-) radical pair were compared in wild-type reaction centers from Rhodobacter sphaeroides and in seven mutants in which the free energy gap, ΔG, between the charge separated states P(+)B(A)(-) and P(+)H(A)(-) was either increased or decreased. Five of the mutant RCs had been described previously, and X-ray crystal structures of two newly constructed complexes were determined by X-ray crystallography. The charge recombination reaction was accelerated in all mutants with a smaller ΔG than in the wild-type, and was slowed in a mutant having a larger ΔG. The free energy difference between the state P(+)H(A)(-) and the PH(A) ground state was unaffected by most of these mutations. These observations were consistent with a model in which the P(+)H(A)(-) â PH(A) charge recombination is thermally activated and occurs via the intermediate state P(+)B(A)(-), with a mean rate related to the size of the ΔG between the states P(+)B(A)(-) and P(+)H(A)(-) and not the ΔG between P(+)H(A)(-) and the ground state. A more detailed analysis of charge recombination in the mutants showed that the kinetics of the reaction were multiexponential, and characterized by ~0.5, ~1-3, and 7-17 ns lifetimes, similar to those measured for wild-type reaction centers. The exact lifetimes and relative amplitudes of the three components were strongly modulated by the mutations. Two models were considered in order to explain the observed multiexponentiality and modulation, involving heterogeneity or relaxation of P(+)H(A)(-) states, with the latter model giving a better fit to the experimental results.
Asunto(s)
Proteínas Bacterianas/metabolismo , Bacterioclorofilas/metabolismo , Feofitinas/metabolismo , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Bacterioclorofilas/química , Bacterioclorofilas/genética , Cristalografía por Rayos X , Transporte de Electrón , Cinética , Modelos Moleculares , Mutación , Feofitinas/química , Feofitinas/genética , Proteínas del Complejo del Centro de Reacción Fotosintética/química , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Rhodobacter sphaeroides/química , Rhodobacter sphaeroides/genéticaRESUMEN
Molecular mass of the iron-dextran complex (M(w)=1133 kDa), diameter of its particles (â¼8.3 nm) and the content of iron ions in the complex core (N(Fe)=6360) were determined by static light scattering, measurements of refractive index increment and the Cotton-Mouton effect in solution. The known number of iron ions permitted the calculation of the permanent magnetic dipole moment value to be µ(Fe)=3.17×10(-18) erg Oe(-1) and the determination of anisotropy of linear magneto-optical polarizabilities components as Δχ=9.2×10(-21) cm(3). Knowing both values and the value of the mean linear optical polarizability α=7.3×10(-20) cm(3), it was possible to show that the total measured CM effect was due to the reorientation of the permanent and the induced magnetic dipole moments of the complex. Analysis of the measured magneto-optical birefringence indicated very small optical anisotropy of linear optical polarizability components, κ(α), which suggested a homogeneous structure of particles of spherical symmetry.
Asunto(s)
Dextranos/química , Hierro/química , Magnetismo , Óptica y Fotónica , Luz , Estructura Molecular , Tamaño de la Partícula , Dispersión de Radiación , Propiedades de SuperficieRESUMEN
Electron transfer at the reaction center of the purple photosynthetic bacterium Rb. sphaeroides R-26 was measured at room temperature by the time-resolved transient absorption spectroscopy technique with 200 fs temporal resolution. The absorbance changes characteristic of the excited state of the primary donor and extending over the whole spectral range investigated from 350 nm up to 720 nm appeared after excitation with a laser pulse of about 100 fs duration at 800 nm. The time evolution of the spectra reflected the excitation of bacteriochlorophylls (BChl) M and L and the subsequent transfer of this excitation to the primary electron donor (P), with the time constant shorter than 1 ps. The decay time constant of the excited primary donor P was determined as about 3 ps, and it was faster than the rise of the reduced intermediary acceptor bacteriopheophytin (BPhe(L)). Photoreduction of BPhe(L) and its further reoxidation was clearly observed as an increase in its bleaching band intensity at around 540 nm in about 4 ps and its decrease in about 200 ps. Our findings support the theoretical model assuming the involvement of the intermediate state P(+)BChl- in the so-called "two-step" model. In this model an electron is transferred in a sequence from the excited special pair P* to bacteriochlorophyll, BChl(L), then to bacteriopheophytin, BPhe(L), and further on to quinone, Q(A). The branched charge separation, partially via P and partially via BChl(L), was also observed.
Asunto(s)
Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Rhodobacter sphaeroides/metabolismo , Absorción , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Transporte de Electrón , Cinética , Fotosíntesis , Proteínas del Complejo del Centro de Reacción Fotosintética/química , EspectrofotometríaRESUMEN
The effect of different ions on the formation and behavior of quadruplex structures of the human telomere sequence d(TTAGGG)(4) has been studied by photon correlation spectroscopy (PCS) and circular dichroism (CD). The saturation and melting curves obtained in the presence of K(+), Na(+), Rb(+), Li(+), Cs(+), and Sr(2+) ions were recorded by CD spectroscopy and indicated the formation of monomeric quadruplexes. Analysis of the saturation curves obtained at 2 degrees C has shown that the presence of a single Sr(2+) ion per oligomer is sufficient for the formation of a monomeric quadruplex of the DNA sequence studied. In the presence of SrCl(2) at a concentration of 50 mM, the formation of tetrameric quadruplexes has been detected. The effect of Sr(2+) ions on the formation of quadruplex structures by the human telomere sequence d(TTAGGG)(4) is stronger and different from that of the other ions tested. The paper also presents results of a study of electrostatic interactions in solution. The translation diffusion coefficients D(T) of the structures present in solution have been determined by photon correlation spectroscopy and the effective charges on the structures have been calculated by combining the experimental data with the results based on the coupled mode theory. Analysis of the melting points monitored by the CD method has permitted a determination of Deltan, the number of ions released in the process of thermal denaturation. All the results are in good agreement with the predictions based on the theory of polyelectrolytes. The effect of ions on the formation and behavior of quadruplex structures of the human telomere sequence d(TTAGGG)(4) has been studied by photon correlation spectroscopy and circular dichroism.
Asunto(s)
Dicroismo Circular/métodos , ADN/química , Iones , Polimorfismo Genético , Análisis Espectral/métodos , Telómero/ultraestructura , Algoritmos , Humanos , Concentración de Iones de Hidrógeno , Modelos Estadísticos , Oligonucleótidos/química , Fotones , Dispersión de Radiación , Electricidad Estática , TermodinámicaRESUMEN
The structure of the Tetrahymena thermophila telomeric sequence d(TGGGGT)(4) was studied by photon correlation spectroscopy (PCS) in aqueous solution in the presence of NaCl, KCl and SrCl(2). The sample studied was polydisperse in all conditions studied. Translational diffusion coefficients D(T) describing the diffusion modes observed were determined. On the basis of a comparison between the experimental D(T) values with those calculated assuming the bead model, two forms were identified as telomeric quadruplex structures: monomer and tetramer. In the presence of SrCl(2) formation of aggregates was observed, with a size that reached several micrometres. The relative weighted concentrations of the structures observed for different concentrations of a salt and DNA were determined. The results obtained in the presence of monovalent ions were qualitatively similar and could be presented in a coherent plot in which the concentration of salt was expressed by the number of ions per DNA molecule. A large number of ions per DNA molecule favoured tetramer formation while a small number favoured the monomer form. A structural phase transition from the monomer to the tetramer induced by a change in the number of ions per DNA molecule was observed. The main difference between the results for Na(+) and K(+) was a greater effectiveness of the K(+) ions in formation of tetramers. The effect of Sr(2+) ions on the structures formed was different than that of the monovalent ions. The results obtained in the presence of Sr(2+) could not be described as a function of the number of ions per DNA molecule.