细胞色素b6f蛋白复合体中叶绿素 a的性质和功能研究

细胞色素b6f蛋白复合体（Cytochrome b6f complex, Cyt b6f）是光合膜上参与光合作用原初反应过程的主要膜蛋白超分子复合体之一。莱茵衣藻和嗜热蓝细菌的Cyt b6f三维晶体结构均显示，每Cyt b6f单体分子含有1分子Chlorophyll a (Chl a )，充分肯定了Chl a 是Cyt b6f天然成分的观点(Kurisu et al，2003;Stroebel et al，2003)。研究表明不同来源的Cyt b6f中Chla单线激发态寿命(或荧光寿命)并不一样，多数的研究结果认为Cyt b6f中Chla单线激发态寿命只有200ps左右，但是也有Cyt b6f中Chla单线激发态寿命为～600ps的报道;而甲醇中游离Chl a 的单线激发态寿命为4ns左右。针对Cyt b6f中Chla单线激发态寿命快速淬灭的现象，Dashdorj 等(2005)根据晶体结构推测Cyt b6f中Chla单线激发态和邻近的Cyt b6亚基上Tyr105残基发生电子交换传递，从而快速淬灭Chla单线激发态，减少了三线态Chl a和单线态氧的产生，并且认为这是Cyt b6f保护自身不受单线态氧破坏的一种机制，但是这一推测缺乏有力的证据。另外，Cyt b6f中Chla的功能仍然未知。本文以菠菜Cyt b6f为对象，结合多种实验手段，测定了菠菜Cyt b6f中Chl a单线激发态寿命，并对复合体中Chl a 单线激发态淬灭的机理进行了深入研究。此外，我们还对复合体中Chl a 可能的功能进行了初步地探讨。获得了如下的结果： 1．针对不同来源的Cyt b6f中Chla单线激发态寿命（或荧光寿命）测定结果不同的报道，仔细分析了其中的原因，发现除了样品来源的差异外，使用不同的去垢剂可能是一个不可忽视的因素。在实验中，不同的研究者分别采用了十二烷基麦芽糖苷（n-Dodecyl β-D-maltoside，DDM，）和八烷基葡萄糖苷（n-Octyl β-D-glucopyranoside，β－OG)作为溶解样品的去垢剂。因此，本文借助稳态吸收和稳态荧光光谱、瞬态光散射技术，CD光谱和亚皮秒时间分辨吸收光谱等技术，分别研究了这两种去垢剂对Cyt b6f结构和功能的不同影响。结果表明，DDM去垢剂能使Cyt b6f处于较好的分散体系中，其中血红素和Chl a分子处于特定的蛋白环境中，不会导致Cyt b6f变性;而β－OG去垢剂会使Cyt b6f产生聚合现象，其中的血红素和Chl a与蛋白环境的相互作用减弱，和DDM相比其电子传递活性显著降低，Chl a单线激发态寿命延长，Chl a更容易被光破坏。通过这一工作，我们优化和确定了Cyt b6f的溶解条件，为下面的研究工作打下了良好的基础。 2．利用Tyr的特异性修饰剂p-Nitrobenzenesulfonyl Fluoride（NBSF）对Cyt b6f样品进行特性修饰，经原子吸收谱、荧光谱、CD谱、质谱等方法对修饰后的样品进行鉴定，并结合时间分辨飞秒吸收光谱技术，测得修饰后的样品在660nm激发下Chl a 单线激发态寿命延长，从而在实验上提供了Tyr与淬灭Chla单线激发态有关的证据。但是对Cyt b6f 中Chl a瞬态吸收图谱仔细研究表明，菠菜Cyt b6f 中Chl a单线激发态快速淬灭过程中并没有发现Tyr与Chl a 之间发生电子传递时所形成的Chla•¯。以上结果表明，Cyt b6f 中Chl a单线激发态快速的淬灭确实和邻近的Tyr105有关，但是与Dashdorj 等提出的Chla单线激发态和Tyr105残基发生了电子交换传递从而淬灭Chla单线激发态这一想法不符，关于这一问题值得进一步深入研究。 3．红光和绿光对Cyt b6f 照射，Cyt f可以被还原，并且红光比绿光更容易使Cyt f 还原，暗示Cyt f 的还原与Chl a 的关系密切，有可能是Chl a 被激发后，其被激发的电子传给Cyt f。对这一现象的进一步研究表明，Cyt b6f在光照条件下，Cyt f 的还原与体系内C10－PQ库的氧化还原状况相关，氧化型的C10－PQ可能介导电子从Chla传向Cyt f。由于在体内质体醌库的氧化还原状态往往决定Cyt b6f 的氧化还原状态，而通过对Cyt b6f不同氧化和还原状态的吸收谱和荧光谱的研究表明，氧化态和还原态的Cyt b6f 中，Chl a 与蛋白的结合状态是有差异的。这些差异可能暗示着Chl a 分子在行使其功能时与复合体的氧化还原状态是有关系的。通过以上的结果，对Cyt b6f中Chl a 可能的功能进行了假设。 4．此外，我们还发现Cyt b6f具有明显的过氧化物酶活性。在0.1 mmol/L乙酸钠缓冲液，pH3.6，25℃，[H2O2] <40mmol/L的条件下，其催化反应的速度常数为kobs＝26±1.2M•s-1;对H2O2的Km 值为50mmol/L，对guaiacol的Km 值为2mmol/L;反应的最适pH为3.6，最适离子强度为0.1，最适温度为35℃。推测Cyt b6f的这种过氧化物酶活性可能是在胁迫环境下使Cyt b6f中的血红素和Chl a 分子免受H2O2的破坏。

Time-Dependent Density Functional Molecular Orbital and Excited State Calculations on Bis(porphyrinyl)butadiynes in the Monocationic, Neutral, Monoanionic, and Dianionic Oxidation States

We report a theoretical study of the multiple oxidation states (1+, 0, 1−, and 2−) of a meso,meso-linked diporphyrin, namely bis[10,15,20-triphenylporphyrinatozinc(II)-5-yl]butadiyne (4), using Time-Dependent Density Functional Theory (TDDFT). The origin of electronic transitions of singlet excited states is discussed in comparison to experimental spectra for the corresponding oxidation states of the close analogue bis{10,15,20-tris[3‘,5‘-di-tert-butylphenyl]porphyrinatozinc(II)-5-yl}butadiyne (3). The latter were measured in previous work under in situ spectroelectrochemical conditions. Excitation energies and orbital compositions of the excited states were obtained for these large delocalized aromatic radicals, which are unique examples of organic mixed-valence systems. The radical cations and anions of butadiyne-bridged diporphyrins such as 3 display characteristic electronic absorption bands in the near-IR region, which have been successfully predicted with use of these computational methods. The radicals are clearly of the “fully delocalized” or Class III type. The key spectral features of the neutral and dianionic states were also reproduced, although due to the large size of these molecules, quantitative agreement of energies with observations is not as good in the blue end of the visible region. The TDDFT calculations are largely in accord with a previous empirical model for the spectra, which was based simplistically on one-electron transitions among the eight key frontier orbitals of the C4 (1,4-butadiyne) linked diporphyrins.

Time-Resolved Resonance Raman Studies on Proton-Induced Electron-Transfer Reaction from Triplet Excited State of 2-Methoxynaphthalene to Decafluorobenzophenone

In this paper time-resolved resonance Raman (TR3) spectra of intermediates generated by proton induced electron-transfer reaction between triplet 2-methoxynaphthalene ((ROMe)-R-3) and decafluorobenzophenone (DFBP) are presented The TR3 vibrational spectra and structure of 2-methoxynaphthalene cation radical (ROMe+) have been analyzed by density functional theory (DFT) calculation It is observed that the structure of naphthalene ring of ROMe+ deviates from the structure of cation radical of naphthalene

Time-resolved resonance Raman spectroscopic studies on the triplet excited state of fluoranil

Perfluoro substituted organic compounds have attracted attention owing to their unique structure and reactivity induced by the perfluoro effect. Fluoranil, a perfluoro derivative of p-benzoquinone, is the subject of this paper. Although the perfluoro effect in the ground state seems to have been well understood there is no information available about such effects on the excited state. Here, the time-resolved resonance Raman spectra of the triplet excited state of fluoranil are reported along with the Raman excitation profiles (REPs) of the various vibrational modes. The vibrational spectral analyses have been carried out by analogy with the fluoranil ground state, triplet benzoquinone, and triplet chloranil vibrational spectral assignments. Also, the assignments are further supported by the calculated frequencies using ab initio theoretical methods. It is observed that for fluoranil in the triplet excited state, due to the perfluoro effect, the structure is considerably less distorted than benzoquinone and also the electron delocalization in the pi* antibonding orbital is less than that of triplet excited state of benzoquinone.

Excited state structure and dynamics of p-benzoquinone and bromanil from time-resolved resonance Raman spectra and simulation

p-Benzoquinone and its halogen substituted derivatives are known to have differing reactivities in the triplet excited state. While bromanil catalyzes the reduction of octaethylporphyrin most efficiently among the halogenated p-benzoquinones, the reaction does not take place in presence of the unsubstituted p-benzoquinone (T. Nakano and Y. Mori, Bull. Chem. Soc. Jpn., 67, 2627 (1994)). Understanding of such differences requires a detailed knowledge of the triplet state structures, normal mode compositions and excited state dynamics. In this paper, we apply a recently presented scheme (M. Puranik, S. Umapathy, J. G. Snijders, and J. Chandrasekhar, J. Chem, Phys., 115, 6106 (2001)) that combines parameters from experiment and computation in a wave packet dynamics simulation to the triplet states of p-benzoquinone and bromanil. The absorption and resonance Raman spectra of both the molecules have been simulated. The normal mode compositions and mode specific excited state displacements have been presented and compared. Time-dependent evolution of the absorption and Raman overlaps for all the observed modes has been discussed in detail. In p-benzoquinone, the initial dynamics is along the C=C stretching and C-H bending modes whereas in bromanil nearly equal displacements are observed along all the stretching coordinates.

Time-Resolved Resonance Raman Spectroscopic Studies on the Triplet Excited State of Thioxanthone

Thioxanthone has been investigated extensively owing to its unique photochemical and photophysical applications and its solvatochromic behavior. Here, we report the time-resolved resonance Raman studies on the structure of the lowest triplet excited state of thioxanthone in carbon tetrachloride. In addition, FT-IR and FT-Raman techniques have been used to study the vibrational structure in the ground state. To corroborate the experimental findings, density functional theory calculations have been carried out. Isotopic calculations and normal coordinate analysis have been used to help in assigning the observed bands to Raman vibrational modes. Structural information derived from this study is expected to help in better understanding the triplet state photochemistry of thioxanthone.

Solvent-induced changes on the polarity of the triplet excited state of 2-chlorothioxanthone: From time-resolved absorption and resonance Raman spectroscopies

Solvent polarity has been known to influence the triplet state structure and reactivity. Here, we present our experimental and theoretical study on the effect of solvent on the lowest triplet excited state structure of 2-chlorothioxanthone (CTX). Time-resolved absorption (TA) spectroscopy has been employed to understand the triplet state electronic structure; whereas solvent-induced structural changes have been studied using time-resolved resonance Raman (TR3) spectroscopy. Both the DFT and TD-DFT calculations have been performed in the solution phase employing self-consistent reaction field implicit solvation model to support the experimental data. It has been observed that CO stretching frequencies of the excited triplet state are sensitive to the solvent polarity and increase with the increase in the solvent polarity. Both TA and TR3 studies reveal that specific solvent effect (H-bonding) is more pronounced in comparison to the nonspecific solvent effect. (C) 2013 Elsevier B.V. All rights reserved.

TIME-RESOLVED ABSORPTION, INFRARED, AND RESONANCE RAMAN-SPECTRA OF THE COMPLEXES [RU(X)(R)(CO)(2)(ALPHA-DIIMINE)] (X=HALIDE R=ALKYL) - INFLUENCE OF X ON THE CHARGE-TRANSFER CHARACTER OF THE LOWEST EXCITED-STATE

Nanosecond time-resolved absorption (TA), resonance Raman (TR(3)), and infrared (TRIR) spectra are reported for several complexes [Ru(X)(R)(CO)(2)(alpha-diimine)] (X = Cl, Br, I; R = Me, Et; alpha-diimine = N,N'-diisopropyl-1,4-diaza-1,3-butadiene (iPr-DAB), pyridine-2-carbaldehyde-N-isopropylimine (iPr-PyCa), 2,2'-bipyridine (bpy)). This is the first instance in which the TA, TR(3), and TRIR techniques have been used to probe excited states in the same series of complexes. The TA spectra of the iodide complexes show a transient absorption between 550 and 700 nm, which does not depend on the solvent but shifts to lower energy in the order iPr-DAB > bpy > iPr-PyCa. This band is assigned to an intraligand transition. For the corresponding chloride and bromide complexes this band occurs at higher energy, most probably because of a change of character of the lowest excited state from XLCT to MLCT. The TRIR spectra show an increase in v(CO) (and k(CO)) on promotion to the excited state; however, the shifts Delta v(CO) show a decrease in the order Cl- > Br- > I-. The TR(3) spectra of the excited complexes [Ru(X)(R)(Co)(2)(iPr-DAB)] show v(s)(CN) of the iPr-DAB ligand 50-80 cm(-1) lower in frequency than for the complexes in their ground state. This frequency shift decreases in the order Cl- > Br- > I-, indicating a decrease of CT character of the lowest excited state in this order. However, going from X = Br to I, the effect on Delta v(CO) is much larger than the decrease of Delta v(s)(CN). This different effect on the CO- and CN-stretching frequencies is assigned to a gradual change in character of the lowest excited state from MLCT to XLCT when Cl- is replaced by Br- and I-. This result confirms a similar conclusion derived from previous resonance Raman and emission experiments on these complexes.

Long-lived excited-state dynamics of i-motif structures probed by time-resolved infrared spectroscopy

UV-generated excited states of cytosine (C) nucleobases are precursors to mutagenic photoproduct formation. The i-motif formed from C-rich sequences is known to exhibit high yields of long-lived excited states following UV absorption. Here the excited states of several i-motif structures have been characterized following 267 nm laser excitation using time-resolved infrared spectroscopy (TRIR). All structures possess a long-lived excited state of ~300 ps and notably in some cases decays greater than 1 ns are observed. These unusually long-lived lifetimes are attributed to the interdigitated DNA structure which prevents direct base stacking overlap.

Time-resolved spectroscopy of the metal-to-metal charge transfer excited state in dinuclear cyano-bridged mixed-valence complexes

Visible pump-probe spectroscopy has been used to identify and characterize short-lived metal-to-metal charge transfer (MMCT) excited states in a group of cyano-bridged mixed-valence complexes of the formula [(LCoNCMII)-N-III(CN)(5)](-), where L is a pentadentate macrocyclic pentaamine (L-14) or triamine-dithiaether (L-14S) and M is Fe or Ru. Nanosecond pump-probe spectroscopy on frozen solutions of [(LCoNCFeII)-Co-14-N-III(CN)(5)](-) and [(LCoNCFeII)-Co-14S-N-III(CN)(5)](-) at 11 K enabled the construction of difference transient absorption spectra that featured a rise in absorbance in the region of 350-400 nm consistent with the generation of the ferricyanide chromophore of the photoexcited complex. The MMCT excited state of the Ru analogue [(LCoNCRuII)-Co-14-N-III(CN)(5)](-) was too short-lived to allow its detection. Femtosecond pump-probe spectroscopy on aqueous solutions of [(LCoNCFeII)-Co-14-N-III(CN)(5)](-) and [(LCoNCFeII)-Co-14S-N-III(CN)(5)](-) at room temperature enabled the lifetimes of their Co-II-Fe-III MMCT excited states to be determined as 0.8 and 1.3 ps, respectively.

Excited-state dynamics in diketopyrrolopyrrole-based copolymer for organic photovoltaics investigated by transient optical spectroscopy

We investigate the photoexcited state dynamics in a donor-acceptor copolymer, poly{3,6-dithiophene-2-yl-2,5-di(2-octyldodecyl)-pyrrolo[3,4-c]- pyrrole-1,4-dione-alt-naphthalene} (pDPP-TNT), by picosecond fluorescence and femtosecond transient absorption spectroscopies. Timeresolved fluorescence lifetime measurements of pDPP-TNT thin films reveal that the lifetime of the singlet excited state is 185 ± 5 ps and that singlet-singlet annihilation occurs at excitation photon densities above 6 × 1017 photons/cm3. From the results of singlet-singlet annihilation analysis, we estimate that the single-singlet annihilation rate constant is (6.0 ± 0.2) × 109cm3 s-1 and the singlet diffusion length is -7 nm. From the comparison of femtosecond transient absorption measurements and picosecond fluorescence measurements, it is found that the time profile of the photobleaching signal in the charge-transfer (CT) absorption band coincides with that of the fluorescence intensity and there is no indication of long-lived species, which clearly suggests that charged species, such as polaron pairs and triplet excitons, are not effectively photogenerated in the neat pDPP-TNT polymer.

Spectral Migration of Fluorescence in Graphene Oxide Aqueous Dispersions: Evidence for Excited-State Proton Transfer

Aqueous dispersions of graphene oxide (GO) exhibit strong pH-dependent fluorescence in the visible that originates, in part, from the oxygenated functionalities present. Here we examine the spectral migration on nanosecond time-scales of the pH dependent features in the fluorescence spectra. We show, from time-resolved emission spectra (TRES) constructed from the wavelength dependent fluorescence decay curves, that the migration is associated with excited state proton transfer. Both `intramolecular' and `intermolecular' transfer involving the quasi-molecular oxygenated aromatic fragments are observed. As a prerequisite to the time-resolved measurements, we have correlated the changes in the steady state fluorescence spectra with the sequence of dissociation events that occur in GO dispersions at different values of pH.

Combined transparent electrodes for high power GaN-based LEDs with long life time

In this paper fabrication of high power light emitting diodes (LEDs) with combined transparent electrodes on both P-GaN and N-GaN have been demonstrated. Simulation and experimental results show that comparing with traditional metal N electrodes the efficacy of LEDs with transparent N electrode is increased by more than 10% and it is easier in process than the other techniques. Further more, combining the transparent electrodes with dielectric anti-reflection film, the extraction efficiency can be improved by 5%. At the same time, the transparent electrodes were protected by the dielectric film and the reliability of LEDs can be improved.

yambo: An ab initio tool for excited state calculations

yambo is an ab initio code for calculating quasiparticle energies and optical properties of electronic systems within the framework of many-body perturbation theory and time-dependent density functional theory. Quasiparticle energies are calculated within the GW approximation for the self-energy. Optical properties are evaluated either by solving the Bethe-Salpeter equation or by using the adiabatic local density approximation. yambo is a plane-wave code that, although particularly suited for calculations of periodic bulk systems, has been applied to a large variety of physical systems. yambo relies on efficient numerical techniques devised to treat systems with reduced dimensionality, or with a large number of degrees of freedom. The code has a user-friendly command-line based interface, flexible 110 procedures and is interfaced to several publicly available density functional ground-state codes.

Ground- and excited-state properties of M-center oxygen vacancy aggregates in the bulk and the surface of MgO

Aggregates of oxygen vacancies (F centers) represent a particular form of point defects in ionic crystals. In this study we have considered the combination of two oxygen vacancies, the M center, in the bulk and on the surface of MgO by means of cluster model calculations. Both neutral and charged forms of the defect M and M+ have been taken into account. The ground state of the M center is characterized by the presence of two doubly occupied impurity levels in the gap of the material; in M+ centers the highest level is singly occupied. For the ground-state properties we used a gradient corrected density functional theory approach. The dipole-allowed singlet-to-singlet and doublet-to-doublet electronic transitions have been determined by means of explicitly correlated multireference second-order perturbation theory calculations. These have been compared with optical transitions determined with the time-dependent density functional theory formalism. The results show that bulk M and M+ centers give rise to intense absorptions at about 4.4 and 4.0 eV, respectively. Another less intense transition at 1.3 eV has also been found for the M+ center. On the surface the transitions occur at 1.6 eV (M+) and 2 eV (M). The results are compared with recently reported electron energy loss spectroscopy spectra on MgO thin films.