ESR STUDIES OF CHLOROPHYLL ONE-ELECTRON PHOTOCHEMISTRY: KINETICS AND MECHANISM OF REVERSIBLE HYDROQUINONE OXIDATION IN SOLUTION*

Abstract— ESR studies have been made of the kinetics of semiquinone radical formation and disappearance resulting from the reversible photosensitization by chlorophyll of hydroquinone oxidation in a pyridine-water solvent. The rate of radical decay was found to be second order with respect to the radical concentration, with a rate constant of 6.7 × 10⁵ l./mole sec at -30°C and an activation energy of 6900 cal/mole. The rate of radical formation was recombination-limited and, through the use of β-carotene as a quencher, the rate constant was determined to be 8.81 × 10⁵ l./mole sec at -30°C. The effect of light intensity and hydroquinone concentration on the rate of semiquinone radical formation and on the steady state radical concentration was also investigated and possible mechanisms to explain the results are discussed.     

FLUORESCENCE SPECTROSCOPY OF A P700-CHLOROPHYLL-PROTEIN COMPLEX*

Abstract. Chlorophyll-protein complexes enriched in the Photosystem I reaction center chlorophyll (P700) exhibit a fluorescence emission maximum at 696 nm at - 196°C The height of this 696 nm emission relative to the emission at 683 nm from antenna chlorophyll a increases proportionally with the P700 concentration while the total fluorescence yield of the complex decreases. The 696 nm emission could possibly be from an absorbing form of antenna chlorophyll a that may be somewhat enriched along with P700 in Photosystem I fractions. However, evidence resulting from glycerol treatment which appears to decrease the rate of resonance energy transfer between antenna chlorophyll and P700 favors the hypothesis that the emission comes from a photooxidized P700 dimer (Chl⁺-Chl) absorbing near 690 nm. In turn, this fluorescence evidence provides additional support for the model of a P700 dimer involving exciton interaction. Absorption in the wavelength region of 450 nm specifically excites emission at 696 nm from the P700-chlorophyll complex.     

Excited state decay of tetrahalomanganese(ii) complexes

The excitation and emission spectra and decay times of several MnX^2-₄ (X = Cl^?, Br^?, 1^?) complexes of various tetraalkylammonium, -phosphonium, and -arsonium salts have been measured for the pure solids at 298°K and 77°K. High luminescence quantum yields (0.3-1.0) reveal that lifetimes fairly accurately reflect radiative decay rates. An impressive correlation exists between the lifetime, τ, of the ⁴T₁ (G) →⁶ A₁ emission and the ligand, X: for X = Cl^?, τ = 1.2 - 3,5 x 10^?3 sec; X = Br^?, τ = 0.35 - 0.43 X 10^?3 sec; X =l^?, τ = 0.036 – 0.055 X 10^?3 sec. We attribute this decreasing lifetime largely to the enhanced spin-orbital coupling associated with the heavier halide. We find that direct population of high energy charge-transfer (CT) states gives smaller emission yields than excitations in the ligand-field (LF) region.     

五氯亚硝酰合铱酸根与二苯胺反应的研究

本文主要用分光光度法研究[Ir(NO)Cl_5]-与二苯胺的反应。在6M盐酸溶液中,生成1:1及2:1配合物,吸收峰分别在515nm及571nm,摩尔吸光系数为2.3×10~4及3.8×10~4。实验表明,新配合物是由[Ir(NO)Cl_5]-的NO~+基在二苯胺氨基的对位发生亲电取代反应而生成。     

PHOTOCHEMISTRY ON SURFACES. EXCITED STATE BEHAVIOR OF RUTHENIUM TRIS(BATHOPHENANTHROLINE DISULFONATE) ON COLLOIDAL ALUMINA-COATED SILICA PARTICLES*

Abstract— The excited state behavior of an anionic ruthenium(II) complex, RuL₃^4-(L=bathophen-anthroline disulfonate), that is electrostatically bound to positively charged alumina-coated silica particles is investigated. The apparent association constant for the binding of RuL₃^4-to the particles is 1.2 × 10⁴M^-1. Surface photochemical processes result in decreased emission yields and multiexponential excited state decay. Excited state quenching by ground-state molecules is evident at high surface coverages. The non-exponential decay kinetics observed at low surface coverages can be attributed either to clustering of the RuL₃^4-molecules or photoionization promoted by Lewis acid sites on the particle surface.     

CORRELATION BETWEEN DELAYED LIGHT EMISSION AND FLUORESCENCE OF CHLOROPHYLL a IN SYSTEM II PARTICLES DERIVED FROM SPINACH CHLOROPLASTS

Abstract— The induction transient of delayed light of chlorophyll a, excited by repetitive flashes (0.5 ms in duration) and emitted 0.1 - 1.2 ms after the flashes, was measured in system II particles derived from spinach chloroplasts. An uncoupler, gramicidin S, was always added to the particles in order to eliminate the influence of the phosphorylation system on the delayed light and to isolate a direct relationship between the delayed light emission and the primary photochemical reaction, except for the experiments described in the next paragraph. The yield of delayed light emission from the system II particles was found to be about three–times higher than that of chloroplasts on a chlorophyll content basis. System I particles, on the other hand, emitted much weaker delayed light than chloroplasts. Upon intermittent illumination, induction of delayed light in system II particles showed a decrease from the initial rise level to the steady-state level. The initial rise level was the maximum. The fluorescence induction, on the other hand, exhibited an increase from the initial rise level to the maximum steady-state level. The induction of both delayed light emission and fluorescence arrived at their final steady-state levels after the same period of illumination. Induction of delayed light emission was measured under various conditions that changed the oxidation-reduction state of the primary electron acceptor, X, of photoreaction II: by adding an electron acceptor and an inhibitor of electron transport, and by changing the light intensity. The state of A'was monitored by measuring the fluorescence yield. The yield of delayed light emission excited by each flash was found to depend on the amount of oxidized form of X present before the flash. To examine the role of the primary electron donor Y of photoreaction II in delayed light emission, effects of electron donors of photoreaction II such as Mn²⁺, hydroquinone and p-phenylenediamine were investigated. These agents were found to markedly decrease the yield of delayed light emission without altering the pattern of its induction. They had little effect on the induction of fluorescence. These findings are interpreted by a mechanism in which transformation of the reaction center from the form (X^-Y⁺) into (X Y) produces a singlet excitation of chlorophyll a that is the source of millisecond delayed light emission. This reaction is probably non–physiological and must be very slow if compared to the transformation of (X^-Y⁺) into (X^-Y). Since the form (X^-Y⁺) is produced when the excitation is transferred to the reaction center in the form (XY), it is expected in this scheme that the yield of delayed light emission should depend on the amount of the form (X Y) present before the excitation flashes. Electron donors stimulate transformation of the reaction center from (X^-Y⁺) into (X^-Y). Since this reaction competes with the process of delayed light emission, electron donors are expected to suppress delayed light emission.     

FLUORESCENCE QUENCHING OF ALLOPHYCOCYANIN AND PHYTOCHROME BY ESTRIOL*

Abstract— At 293 K the long-wavelength absorption and emission band of 1.4 μM allophycocyanin is decreased by estriol (Δ¹-³-⁵⁽¹⁰⁾-estratriene-3,16α, 17β-trio!) in the range 0.8-6.6 μM in the presence of 11% alcohol (vol/vol). The binding of estriol is shown to be of high affinity, 1:1 with allophycocyanin. The free energy of this binding process (ΔG^°) is -33.6 kJ mol' and single binding site dissociation constant (K_D) 1.0 ×10–6M. Estriol at 21 μM effectively quenches the fluorescence of 1.4 M large molecular weight phytochrome in its red absorbing form at 77 K while having little or no effect on the phototransformation difference spectrum at 293 K.     

A field ionization study of octan-2-one

H/D randomization is not observed within the octan-2-one-1,1,1,3,3-d₅ molecular ion at times less than 7 × 10^?10 sec following field ionization. Partial H/D randomization is observbed at 10^?6 to 10^?5 sec. It is deduced that the curves of microscopic rate constant k against internal excitation energy E for the reactions effecting randomization and for the McLafferty rearrangement intersect. The field ionization kinetics (FIK) for the McLafferty rearrangement in octan-2-one-1,1,1,3,3-d₅ are investigated over a time range extending from 10^?11 to 10^?6 sec. The maximum microscopic rate constant k for the reaction is estimated as 5 × 10¹⁰ sec^?1. It is suggested that FIK measurements with a double focusing mass spectrometer be made by sweeping the blade potential rather than by the alternative technique of sweeping the electric sector analyzer potential. The ‘normnal’ FI mass spectra of octan-2 one and octan-2-one and octan-2-one-1,1,1,3,3-d₅ are presented and discussed. It is proposed that the m/e 56 species in the ‘normal’ FI mass spectrum of octan-2-one represents a doubly charged ion formed by loss of oxygen in a surface process.     

ESR studies of chlorophyll one-electron photochemistry in solution: kinetics of reversible quinone reduction and general redox mechanisms

Abstract— ESR studies have been made of the kinetics of semiquinone radical formation and disappearance resulting from the reversible photosensitization of reduction or oxidation, by chlorophyll, pheophytin or hematoporphyrin, of several quinone-hydroquinone pairs in various solvents. The rate of radical decay was found to be second order with respect to the radical concentration in all systems. Radical formation rates were determined by the initial production rate minus the decay rate. The kinetic constants for single electron transfer between triplet porphyrins and quinones or hydroquinones were determined usingβ-carotene as a quencher in aqueous pyridine, and by measuring the initial rate of radical formation at various concentrations of quinones and hydroquinones in methanol and ethanol. These constants were found to be approximately the same in a given solvent for benzoquinone and hydroquinone with all porphyrins, though the rates differed in different solvents: pyridine-water ～ 10⁶I./mole sec, and methanol and ethanol ～ 5X 10⁴l./mole sec. Trimethylquinone and its hydroquinone also give similar rate constants for radical formation in pyridine-water, ～ 10⁶ l./mole sec. The second order radical decay constants for both benzoquinone and hydroquinone in pyridine-water were the same, ～ 10⁵ I./mole sec, with either chlorophyll, pheophytin or hematoporphyrin as sensitizer. The same activation energy, 6900 cal/mole, was found for chlorophyll-benzoquinone and hydroquinone in aqueous pyridine; 5500 cal/mole was obtained for these systems in ethanol. In methanol and ethanol solutions of chlorophyll, the same radical decay rate constants, ～10⁶ I./mole sec, were observed for both benzoquinone and hydroquinone. Also, the same decay constants, ～ 10⁶ I./mole sec, were found for trimethylquinone and its hydroquinine in pyridine-water. These latter two compounds gave extremely small steady-state ESR signals in ethanol compared with aqueous pyridine. We have also observed that the steady-state signal obtained with chlorophyll-menadione in ethanol-water was much enhanced by the presence of NADH. In contrast, NAD⁺ was found to decrease radical production, by increasing the decay rate, in the chlorophyll-hydroquinone system in aqueous pyridine. These results are discussed in terms of possible mechanisms for radical formation and disappearance. The most likely possibility is considered to be a one-electron oxidation or reduction of the porphyrin triplet, followed by radical disproportionation and redox reactions between the disproportionation products.     

ANALYSIS OF MICROSECOND FLUORESCENCE YIELD AND DELAYED LIGHT EMISSION CHANGES AFTER A SINGLE FLASH IN PEA CHLOROPLASTS: EFFECTS OF MONO- AND DIVALENT CATIONS

Abstract. New results are presented on the effects of mono- and divalent cations on concurrent changes in the microsecond yields and kinetics of chlorophyll a fluorescence and delayed light emission, and the light saturation curve for the latter at 100 μs, following a 10 ns flash at 337 nm. (1) The fluorescence yield increases exponentially from 3 to 30 μs (lifetime, τ, 6.4 ± 0.6/μs), and decays biphasically between 50 and 800μs. (2) The delayed light emission decays biphasically with two exponential phases: fast phase, T= 7–10μs, and slow phase, T= 33–40μs. (3) The light saturation curve for 100μs delayed light emission is satisfactorily represented by a one-hit Poisson saturation curve. (4) Addition of 5 mM NaCl to salt-depleted chloroplasts decreases (by as much as 40%) the yields of μs fluorescence and delayed light emission, and the subsequent addition of 5mM MgCl₂ increases the yields (≤2 × over samples with only NaCl). (5) The fluorescence yield rise and delayed light emission decay kinetics are independent of low concentrations of cations. The lifetime of the fast phase of fluorescence decay changes from ?90μs to ?160μs, when Na⁺ or Na⁺+ Mg²⁺ are added. Based on a detailed analysis presented in this paper, the following conclusions regarding the effects of low concentrations (few mM) of mono-and divalent cations in sucrose-washed chloroplasts at room temperature are made: (a) Na⁺ decreases (?6%) and Mg²⁺ increases (? 20% compared with the Na⁺ sample) the sensitization of photosystem II photochemistry: this effect is small, but significant. (b) Na⁺ increases and Mg²⁺ decreases the efficiency for radiationless transitions in singlet excited Chl a in the antenna and closed reaction center of PS II; this includes non-radiative energy transfer to PS I, intramolecular intersystem crossing and internal conversion. The ratio of the sum of the rate constants for radiationless transitions to that for fluorescence increases by ? 2-fold upon the addition of Na⁺, and is completely reversed by the addition of Mg²⁺. (c) The rate constant for the re-oxidation of Q^- decreases (about 50%) in the presence of Na⁺ or Na⁺+ Mg²⁺. These conclusions imply that cations produce multiple changes in the primary photoprocesses of PS II at physiological temperatures. It is proposed that these changes are mutually independent and can co-exist.     

LASER PHOTOLYSIS STUDIES OF QUINONE REDUCTION BY CHLOROPHYLL a IN ALCOHOL SOLUTION

Upon laser photolysis of chlorophyll-quinone solutions in ethanol, transients due to the chlorophyll triplet state (C_t), the chlorophyll cation radical (C⁺) and the semiquinone radical (Q^-) can be observed. The rise of Q^- parallels the decay of C_t. demonstrating the precursor role of the triplet. The decay of C⁺ is second order, consistent with reverse electron transfer, and has a rate constant which is independent of quinone potential, and an activation energy of 14kJ/mol due mainly to the temperature dependence of solvent viscosity. Triplet quenching and C⁺ yield are found to decrease with decreasing quinone potential.     

Preparation and Characterization of Storage and Emission Functional Material of Chlorine Anion: Ca24Al28O644+·(Cl-)3.80(O2-)0.10

A storage and emission functional material of Ca₂₄Al₂₈O₆₄⁴⁺·(Cl^-)_3.80(O^2-)_0.10(C12A7-Cl^-), was prepared by the solid-state reactions of CaCO₃, γ-Al₂O₃, and CaCl₂ in Cl₂/Ar mixture atmosphere. The anionic species stored in the C12A7-Cl^- material were dominated by Cl^-, about (2.21±0.24)×10²¹ cm^-3, accompanied by a small amount of O^2-, O^-,, and O^2-, measured via ion chromatography, electron paramagnetic resonance, and raman spectra measurements. These results also corroborate identification of time-of-flight mass spectroscopy—the anionic species emitted from the C12A7-Cl^- surface were dominated by the Cl^- (about 90%) together with a small amount of O^-and electrons. The structure and morphological alterations of the material were investigated via X-ray diffraction and field emission scanning electron microscope, respectively.     

钙镁磷肥(FCMP)阴离子构型的硅烷化气相色谱(TMS-GC)研究

通过对钙镁磷肥(FCMP)硅烷化气相色谱(TMS-GC)的研究证明:FCMP中硅以一聚、二聚、三聚链及环四构型为主,同时还有少量其他一维、二维、三维构型存在;而磷则以单个四面体与硅聚合或独立存在。     

Thermoluminescence and temperature effects on delayed light emission (corrected for changes in quantum yield of fluorescence) in DCMU-treated algae

Abstract—

• 1 The simultaneous measurements of delayed light emission (DLE) and chlorophyll (Chl) fluorescence yield in DCMU§ treated Chlorella were made in the time range of 1 to 10 sec at various temperatures from 0 to 50°C. Similar measurements were made for DCMU treated thermophilic strain of Synechococcus in the temperature range of 0 to 75°C.
• 2 Using the basic assumption that DLE is produced by the back reaction of primary photoproducts of system II, and that two such reactions are required for it, a linear relationship between J^-1/2 (where J is energy per unit time available for DLE) and time after illumination was derived. This second-order relationship was confirmed experimentally at several temperatures (2°, 5°, 10° and 15°C). From these analyses, reaction rate decay constants, at specific temperatures, were calculated.
• 3 An Arrhenius plot was made for these calculated rate constants. Its slope (8–10 kcal/mole) agreed well with previous reports; however, it had a region of zero slope which occurred at the physiological temperature of the organisms used.
• 4 Thermoluminescence or temperature jump delayed light emission (TDLE) was measured using various temperature conditions and it was found that not only the magnitude of the temperature jump (ΔT), but the initial and final temperatures of the sample were important. For example, a temperature jump of 8°C from 2 to 10°C gave much higher TDLE than from 12 to 20°C.
• 5 Many properties e.g., magnitude, temperature dependence and time independence of TDLE could be explained by the DLE decay data (corrected for changes in fluorescence yield) and the kinetic analysis.
• 6 It is suggested that, in addition to the back reaction of Z⁺ (the primary oxidized photoproduct of system II) with Q^- (the primary reduced photoproduct of system II), a reducing entity, beyond the sites of DCMU and antimycin a action, is somehow involved in the production of slow DLE.

     

Investigation of the reaction of Br 2 - and Fe(II) using laser pulsed photolysis

Conclusions The reaction of Br ₂ ^- with Fe _solv ²⁺ in a mixture of methanol with diethyl ether occurs according to a intrasphere mechanism with the primary product being the FeBr²⁺ complex. The rate constant (5.9 ± 0.5)-10⁷ dm³/mole·sec at 298 K is greater by a factor of 15–20 than in the reaction of Br ₂ ^- with Fe _aq ²⁺ , despite the slower rate of exchange of the solvent molecules for Fe ₂ ^- . In the latter case the reduced permittivity considerably increases the lifetime of partners in the contact pair state.Translated from Izvestiya Akademii Nauk SSSR, Seriya Khimicheskaya, No. 6, pp. 1247–1250, June, 1988.     

STRUCTURE OF BACTERIORHODOPSIN and in situ ISOMERIZATION OF RETINAL: A MOLECULAR DYNAMICS STUDY*

Abstract– Henderson's model of the structure of bacteriorhodopsin has been completed by adding the missing loop regions and by subsequent energy minimization and equilibration (for about 100 ps) at 300 K. Analysis of the structure during a later 20 ps molecular dynamics run showed no significant deviations from the Henderson model. In situ isomerization reactions of the retinal chromophore in bacteriorhodopsin have then been simulated to investigate the chromophore protein interaction for the three isomerization reactions: (i) all-trans→ 13-cis; (ii)all-trans→ 13,14-dicis; and (iii) all-trans→ 13,15-dicis. We find that reaction (iii) which accompanies dark-adaptation of bacteriorhodopsin can proceed in the binding site without any sterical hinderance and involves negligible motions of the covalently bound Lys-216 and other side groups. Reaction (ii) exhibits a somewhat larger but still small energy barrier and involves little rearrangement of Lys-216 and the protein backbone. Reaction (i) experiences a sterical impediment amounting to more than 10 kT at physiological temperatures and also induces significant structural changes at the binding site. Our simulations also reveal that reaction (ii) as a photo-isomerization process can be completed within about 400 fs, whereas reaction (i) requires longer times for completion. Reaction (i) is also accompanied by a co-rotation of the 14–15 bond by 150° (even when a torsional barrier of 20 kcal/mol is imposed to impede rotation of the 14–15 bond) such that photoreactions (i) and (ii), in effect, lead to very similar final geometries. Isomerization (ii) can readily explain the pump mechanism of bacteriorhodopsin: the sequential, thermal back-reaction 13,14-dicis→ 13-cisall-trans can be acid-base catalyzed, i.e., coupled to deprotonation and reprotonation of retinal's Schiff base nitrogen. The orientation of retinal is such that Asp-85 can act as the acceptor and Asp-96 as the (indirect) donor. The thermal back-reaction 13,14-dicis→ all-trans can be coupled to vectorial Cl^? ion transport as well.     

CHLOROPHYLL a FLUORESCENCE OF GONYAULAX POLYEDRA GROWN ON A LIGHT-DARK CYCLE AND AFTER TRANSFER TO CONSTANT LIGHT

Abstract— The chlorophyll a fluorescence properties of Gonyaulax polyedra cells before and after transfer from a lightdark cycle (LD) to constant dim light (LL) were investigated. The latter display a faster fluorescence transient from the level ‘I’ (intermediary peak) to ‘D’ (dip) to ‘P’ (peak) than the former (3 s as compared to 10 s), and a different pattern of decline in fluorescence from ‘I’ to ‘D’ and from ‘P’ to the steady state level with no clearly separable second wave of slow fluorescence change, referred to as ‘s' (quasi steady state)→‘M’ (maximum) →‘T’ (terminal steady state). The above differences are constant features of cells in LD and LL, and are not dependent on the time of day. They are interpreted as evidence for a greater ratio of photosystem II/photosystem I activity in cells in LL. After an initial photoadaptive response following transfer from LD to LL, the cell absorbance at room temperature and fluorescence emission spectra at 77 K for cells in LL and LD are comparable. The major emission peak is at 685–688 nm (from an antenna Chl a 680, perhaps Chl a-c complex), but, unlike higher plants and other algae, the emission bands at 696–698 nm (from Chl a_II complex, Chl a 685, close to reaction center II) and 710–720 nm (from Chl a₁, complexes, Chl a 695, close to reaction center I) are very minor and could be observed only in the fluorescence emission difference spectra of LL minus LD cells and in the ratio spectra of DCMU-treated to non-treated cells. Comparison of emission spectra of cells in LL and LD suggested that, in LL, there is a slightly greater net excitation energy transfer from the light-harvesting peridinin-Chl a (Chl a 670) complex, fluorescing at 675 nm, to the other antenna chlorophyll a complex fluorescing at 685–688 nm, and from the Chl a., complex to the reaction center II. Comparison of excitation spectra of fluorescence of LL and LD cells, in the presence of DCMU, confirmed that cells in LL transfer energy more extensively from the peridinin-Chl a complex to other Chl a complexes than do cells in LD.     

Structure and spectral property of 8-(2-(5-(4-methylphenyl)-2-thienyl)vinyl)-10,10-dimethyl-10H-pyrido[1,2-a]indolium perchlorate

The title compound 8-(2-(5-(4-methylphenyl)-2-thienyl)vinyl)l-10,10-dimethyl-10H-pyrido[1,2-a]indolium perchlorate (3) has been synthesized by cycloaddition and condensation reactions, and characterized by elemental analysis, IR, ¹H NMR, and single-crystal X-ray diffraction. The crystal structure analysis exhibits that the molecule of 3 possesses good coplanarity, and the three rings (10H-pyrido[1,2-a]indolium, thienyl and phenyl) and vinyl moiety can make up a large conjugated system. Its UV-Vis and fluorescence spectra were measured, and found that it displays larger maximum absorptions and emission wavelengths in comparison with 8-(4-methylphenyl)vinyl analogue.     

LIGHT REQUIREMENTS FOR THE ENHANCED SYNTHESIS OF A PLASTID mRNA DURING SPIRODELA GREENING

A plastid mRNA (5 × 10⁵ mol wt) appears as a burst 3 h after white light greening of steady state dark grown plants of Spirodela oligorrhiza. In this species, chlorophyll synthesis begins after 12 h. The light requirement is different from the pulse of far-red reversible red light required to abolish the lag of chlorophyll synthesis in many species, including Spirodela. Continuous high energy far-red is not stimulatory. When the illumination is not continued throughout the time of incorporation, the stimulation is minimal. Low energy blue and red light are stimulatory, and green and far-red light are ineffectual. Blue light was > 5 times as effective as red light at many dose levels. Illumination with 3 × 10¹⁷ quanta/m²/s (50pEm/cm²/s) blue light at 476 nm gave about half maximum stimulation.     

四氢呋喃开环聚合的研究——Ⅲ.本体聚合的总生长反应速度常数

用溴丙烯-高氯酸银或癸二酰氯-高氯酸银引发四氢呋喃在0℃进行本体聚合,测得总生长反应速度常数(k_p)分别为0.32×10~(-3)及0.38×10~(-3)升·克分子~(-1)·秒~(-1)。这样低的k_p值认为是由于ClO_4~-抗衡负离子具有强的亲核性所致。在溴丙烯-高氯酸银催化体系配此中加大银盐用量(到过量51％),引发诱导时间缩短,聚合速度加快,而k_p值的增大不明显。