HETEROGENEITY OF THE PHOTOCHEMICAL CENTERS IN SYSTEM II OF CHLOROPLASTS*

Abstract— In 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) poisoned chloroplasts of algae and‘ higher plants the area over the fluorescence induction curve increases with biphasic first order kinetics (Melis and Homann, 1975). Two possibilities are considered to explain the biphasic nature of the area growth. The first is a sequential double reduction of the primary electron acceptor in system II while the second envisages a heterogeneity of its photochemical centers. The kinetic properties of the area growth after firing a single saturating flash proved to be incompatible with the predictions of the “sequential double reduction” model. This conclusion was corroborated by results obtained from a kinetic analysis of the area restoration process in the dark, and an analysis of the partially restored areas. Assuming an existence of a heterogeneous pool of photochemical centers, the growth of the area over the fluorescence curve could be further analyzed to yield two components, a fast a-component, and a relatively slow β-component. The kinetic characteristics of these components, and the effect of a short saturating flash on their respective size, led to the conclusion that one type of photochemical center had a faster recombination rate of the photochemically separated charges and was less efficient in trapping excitation energy.     

PAIRWISE RECOMBINATION IN THE DECAY OF PHOTOCHEMICAL HYDRATED ELECTRONS*

Abstract— The decays of hydrated electrons generated by 265 nm laser flash photolysis of aqueous I^-, tyrosine, tryptophan, lysozyrne. and ribonuclease have been analyzed by numerical computing. Homogeneous reactions do not explain the data without e^- _aq—radical back reactions significantly faster than diffusion limited. The results are explained by postulating pairwise recombination persists at times where conventional diffusion theory predicts randomization, particularly at low intensities and in the absence of effective e^- _aq scavengers. The physical mechanisms responsible for these effects are considered.     

REACTION OF EXCITED CHLOROPHYLL MOLECULES AT ELECTRODES AND IN PHOTOSYNTHESIS*

Abstract— Excited molecules can exchange electrons with suitable electrodes in an electrochemical cell. Excitation energy of molecules can therefore be converted into electrochemical energy. Electrochemical reactions of excited chlorophyll molecules have been investigated with the help of semiconductor electrodes. It is suggested that this type of reaction also occurs during the primary steps of photosynthesis. Consequently, concepts of electrochemical kinetics would have to be applied in order to elucidate chlorophyll-sensitized reactions in photosynthetic membranes. In order to provide evidence for this conclusion, electrochemical kinetics is applied to the calculation of decay of delayed light from photosynthetic membranes, and the result is compared with experimental data from Chlorella pyrenoidosa. The conversion of light into electrochemical energy via photoelectrochemical reactions of organic dyes in an electrochemical cell is demonstrated, and the postulated analogous electrochemical mechanism for photosynthesis is discussed.     

PHOTOCHEMICAL INACTIVATION OF SINGLE-STRANDED VIRAL DNA IN THE PRESENCE OF UROCANIC ACID*

Abstract— Urocanic acid (UA) has previously been shown to react photochemically in vitro with N,N-dimethylthymine. In this study, mixtures of UA and phage G4 single-stranded DNA have been irradiated with UV light (λ≥ 254 nm) and the DNA assayed for infectivity. At the concentrations of UA employed (typically 5.4 × 10^-3 M ) there is extensive absorption of the incident light by the UA. The DNA is inactivated at rates greater than that predicted from the calculated shielding by UA, indicating that photosensitization is occurring. Photosensitization is also indicated by the fact that at high UA concentrations the inactivation rate does not decrease to zero but approaches a residual value. Furthermore, the ability to photoreactivate DNA that has been photolyzed in the presence of UA is much reduced relative to that observed upon photolysis of the DNA alone. UA is therefore responsible for the production of UV-induced DNA lesions, which are resistant to photoreactivation.
A general analysis of the effects of photosensitization on the kinetics of UV inactivation is presented in an appendix.     

PHOTOCHEMICAL CHARACTERIZATION OF COVALENTLY-LINKED PORPHYRIN-QUINONE COMPLEXES*

Abstract— The fluorescence properties of a covalently-linked porphyrin-quinone complex and its zinc derivative were studied in a variety of organic solvents. The kinetics of fluorescence decay for both the quinone and hydroquinone oxidation states were measured in acetonitrile, dichloromethane, dimethyl-formamide, and pentane. The fluorescence yield and kinetics of decay at room temperature were little affected in the porphyrin or zinc porphyrin complexes when the attached quinone was reduced. However, for these complexes the fluorescence yield and lifetimes were both substantially decreased in acetonitrile and dichloromethane when the quinone was in its oxidized state. These latter decay kinetics were not explainable by a process having a single exponential decay. On the other hand, little fluorescence quenching or lifetime shortening was observed in dimethylformamide or pentane, indicating unique solvent dependencies for the quenching process. Evidence was obtained for photoproduced charge separation from EPR measurements on the covalently-linked zinc porphyrin-quinone complex. The EPR data showed equivalent concentrations of a Zn porphyrin cation radical and a benzoquinone anion radical in acetonitrile or dichloromethane at both room temperature and 77 K. The charge separated state rapidly decayed at room temperature (in sub-millisecond times) but was quite stable at 77 K. It is concluded that light-induced charge separation in acetonitrile and dichloromethane at room temperature may occur from the excited singlet state with a high quantum efficiency. A photoproduced charge separated state also occurred when the covalently-linked complexes were incorporated into egg yolk phosphatidylcholine liposomes. The quantum yield for radical formation in this latter system was 0.1 and the lifetimes of the radical species formed were many minutes.     

VARIATION IN THE PHOTOCHEMICAL REACTIVITY OF THYMINE IN THE DNA OF B. SUBTILIS SPORES, VEGETATIVE CELLS AND SPORES GERMINATED IN CHLORAMPHENICOL*,†

Abstract— The chief photoproduct of thymine produced in u.v. irradiated (2537Å) vegetative cells of B. subtilis is the cyclobutane-type dimer while in spores very little of this dimer is produced (maximum yield 2·6 per cent of thymine) but a new photoproduct is produced in high yield (maximum of 28·4 per cent of thymine). This difference in photochemical response appears to be due, at least in part, to a difference in uydration of the DNA. The photochemistry of thymine in isolated DNA irradiated in solution is similar to that of DNA in irradiated vegetative cells, but differs markedly from that of isolated DNA irradiated dry. The yield of cyclobutane-type thymine dimer is much reduced in isolated DNA irradiated dry but a new photoproduct of thymine. is produced which is chromatographically similar to the spore photoproduct. The yield of this photoproduct, however, is never as great as that obtained in irradiated spores. The photochemistry of the DNA thymine of spores germinated in the presence of chloramphenicol is very similar to that of normal vegetative cells. Except for hydration, the physical state of the DNA is probably not otherwise altered by germination in the presence of chloramphenicol since DNA replication is prevented by the presence of chloramphenicol. These results are also consistent with the hypothesis that the unique photochemistry of spores is due, at least in part, to the hydration state of the DNA. The acid stability of the spore photoproduct is indicated by the fact that it is isolated from irradiated spores after hydrolysis in trifluoroacetic acid at 155°C for 60 min. It still contains the methyl group of thymine as judged by the fact that for a given dose of u.v. the same yield of photoproduct was obtained whether the spores were labeled with thymine-2–C-14 or -methyl-C-14. This photoproduct is stable to reirradiation (2537Å) in solution under condiditions where thymine dimers of the cyclobutane-type are completely converted back to monomeric thymine. On a column of molecular sieve material (Sephadex-G10), the spore photoproduct elutes in a region intermediate between the cyclobutanetype thymine dimers and monomeric thymine. Of the numerous compounds tested by paper chromatography, the spore photoproduct is most similar (but not identical) in several solvents to 5–hydroxyuracil and 5–hydroxymethyluracil. Our data do not allow us to decide if the product is a monomer or a dimer. Although the photochemistry of thymine in the DNA of spores differs markedly from that for vegetative cells, several lines of evidence make it seem doubtful that the enhanced resistance of spores to u.v. relative to that of vegetative cells can be explained solely on the basis of this difference in the photochemistry of DNA thymine.     

PACKING OF IONS IN SILICATE GLASSES

Any silicate glass can be regarded as the relaxation of SiO_2 glass and the volumeis composed of two parts. One is the topological volume of O~(2-), which is equal to thevolume per oxygen of SiO_2 glass, the other is the topological volumes of cations. Thetopological volume of a monovalent cation equals the volume shared by each sphere indense haphazard packing of equal spheres of the cation's size. The charges of bivalentcations cause a decrease of volume, the decrement being the packing volume of Ca~(2+). Anequation is reached for the calculation of the volume per oxygen of silicate glasses. Alot of calculations show that the equation is accurate, so accurate that the densities canbe calculated. The study reveals some characters of the glass structure and clearly ex-presses the contributions of various cations to the volumes of glasses.     

ELECTROCHEMISTRY OF EXCITED MOLECULES: PHOTO-ELECTROCHEMICAL REACTIONS OF CHLOROPHYLLS*

Abstract— Semiconductors with a sufficiently large energy gap, in contact with an electrolyte, can be used as electrodes for the study of electrochemical reactions of excited molecules. The behavior of excited chlorophyll molecules at single crystal ZnO-electrodes has been investigated. These molecules inject electrons from excited levels into the conduction band of the electrode, thus giving rise to an anodic photocurrent. The influence of various agents on this electron transfer has been studied. In the presence of suitable electron donors (e.g., hydroquinone, phenylhydrazine) in the electrolyte chlorophyll molecules, absorbing quanta, mediate the pumping of electrons from levels of the reducing agents into the conduction band of the semiconductor-electron acceptor. The electron capture by the semiconductor electrode is irreversible, when an adequate electrochemical gradient is provided in the electrode surface. Some properties of excited chlorophyll at semiconductor electrodes (unidirectional electron transfer, highly efficient charge separation, chlorophyll as electron pump and able to convert electronic excitation into electric energy) show similarity to the behavior of chlorophyll in photosynthetic reaction centers.     

ON THE FORMATION AND REACTIVITY OF DIOXIRANE INTERMEDIATES IN THE REACTION OF PEROXOANIONS WITH ORGANIC SUBSTRATES*

Abstract— Kinetics and ¹⁸O-labeling studies have provided evidence for the involvement of dioxirane intermediates (V) in the ketone-catalysed decomposition of peroxomonosulfate (Caroate) HSO;. Reaction rates depend on ketone structure. In competition with catalysis of peroxide decomposition, the dioxirane intermediate is capable of oxidizing several organic and inorganic substrates. Thus, cis- and rrans-cinnammic acids could be converted stereospecifically into the corresponding epoxides. Also, oxidation of phenylpropiolic acid, a substrate which is representative of weakly nucleophilic alkynes, could be carried out using the Caroatehetone oxidizing system. Under the conditions adopted, no oxidation of the substrates examined was found to occur in the absence of ketone. The possibility that formation of dioxathiirane intermediates (XXII) occurs following a side pathway in the reaction of Caroate with sulfoxides (which produces sulfones in high yield) has been explored. Preliminary experiments using ¹⁸O-labeling of p-tolyl phenylsulfoxide, however. failed to support this hypothesis. pointing out the need for further detailed studies.     

PHOSPHORESCENCE OF AROMATIC KETONES IN LOW-TEMPERATURE GLASSES

Abstract— Phosphorescence decay kinetics and emission spectra of some acetophenone derivatives were measured in ether/alcohol and hydrocarbon glasses. Most decay rates observed were a sum of exponentials, and different emission spectra were observed at different intervals in the decay. The observations can be explained in terms of coexistent n , π*, π, π*, and charge-transfer triplets, but solvent shifts in the direction expected for π, π* and charge-transfer triplets were not observed.     

ANALYSIS OF PRIMARY PHOTOCHEMICAL PROCESSES IN BACTERIORHODOPSIN

Abstract— The sequence of primary events following light absorption by light adapted bacteriorhodopsin (bR₅₇₀) is considered by analyzing recent picosecond absorption and emission data. The analysis is facilitated by theoretical calculations which allow us to characterize the properties of the first excited singlet state. It is concluded that excitation leads to the eventual population of a photochemically important nonfluorescent excited state (I) which decays into a photoproduct (J₆₂₅)- In J₆₂₅, which is most probably a ground state molecule, the chromophore has undergone a structural change, presumably trans → 13- cis isomerization. It is suggested that the subsequent process

reflects a relaxation of the protein environment involving proton transfer.     

REACTIVITY OF MACROMONOMERS IN FREE RADICAL POLYMERIZATION

Abstract

Macromonomers are linear polymeric o r oligomeric species which, because of the presence of a reactive end group, have the potential either to polymerize with themselves or with comonomers. The reactive group is most commonly a vinyl group that can participate in free radical polymerization, but any polymerizable end group, such as epoxy, bis-hydroxy, etc., is sufficient for the molecule to be classified as a macromonomer. The words “macromer” and “macromonomer” are often used interchangeably, although the former term was introduced originally as a trademark of CPC International to describe the macromonomers discovered by Milkovich [1].     

PHOTOCHEMICAL ADDITION OF L-LYSINE TO 1,3-DIMETHYL-4-THIOURACIL*

Irradiation of an aqueous solution of 1,3-dimethyl-4-thiouracil (1) in the presence of l -lysine ( 2 ) with a high-presure mercury lamp through a Pyrex filter gave a 1:1 mixture of two photoproducts. One of the products was isolated in a pure form, which on treatment with acetic anhydride-pyridine yielded the starting material 1. Based on the spectral data (¹H and ¹³C NMR) and the chemical reaction, the structure of the photoproduct was assigned to 3. The photochemical reaction is considered as a nucleophilic addition of l -lysine to photoexcited 1.     

PHOTOCHEMICAL DECOMPOSITION OF 1, 4-BENZODIAZEPINES. NITRAZEPAM*

Abstract— The photochemical activity of nitrazepam, a drug of which phototoxic effects are known, is investigated. Nitrazepam decomposes photochemically in an oxygen-poor medium, while it is relatively stable in the presence of oxygen. It appears that this quenching of excited nitrazepam by molecular oxygen leads to the formation of singlet oxygen. This is demonstrated in three different ways: 1. In the presence of 2-methyl-2-pentene as acceptor of singlet oxygen, two characteristic products are formed: 2-methyl-1-penten-3-ol and 2-methyl-3-penten-2-ol. 2. The photosensitized oxidation of L-dopa in the presence of nitrazepam is increased when D₂O, instead of H₂O, is added to the solvent system, due to the longer lifetime of singlet oxygen in D₂O. 3. The β-value, the ratio of the rate of decay of singlet oxygen to its rate of reaction, is determined with L-dopa as acceptor of singlet oxygen. The same value is obtained with both nitrazepam and methylene blue, a known singlet oxygen generator.     

ENERGETICS OF PHOTOPHYSICAL PROCESSES IN CHLOROPHYLL-LIKE MOLECULES

Abstract— The values of the absolute quantum yields of fluorescence and of intersystem crossing have been obtained for porphin and its derivatives by methods based on relative measurements involving flash photolysis. For a given compound the sum of these quantum yields is unity within experimental error which shows that the energetics of the molecules investigated is determined by competition of only two processes, viz., fluorescence and intersystem crossing, while internal conversion plays a negligible role. The values of the corresponding transition probabilities have been determined with the help of phase-fluorometric measurements. The dependence of the transition probabilities on molecular structure is discussed.     

PHOTOCHEMICAL BEHAVIOR OF BACTERIORHODOPSIN IMMOBILIZED IN NaCl PELLETS

Abstract— Some photochemical reactions of bacteriorhodopsin (BR) embedded in NaCl pellets (BR-NaCI) in the visible region are described here. BR in these preparations is a mixture of two classes of species: a drastically blue-shifted form and the unchanged purple pigment. Depending on the illumination history of the BR before being immobilized, both kinds of BR could be demonstrated in light-adapted (LA) and dark-adapted (DA) forms, but light adaptation was not possible once the pellets were made. Analogously to BR suspensions, the light-adapted blue-shifted BRexhibited an a/l-trans type photocycle, but the thermal steps were greatly slowed down (time constants 1 to 5 min). The parent species absorb at 506 nm. The DA blue-shifted BR exhibited absorption changes resolved into two photoreactions, one all-(rans- like (as in LA-BR) and another, 13-cw like, whose decay rate is also greatly slowed down (recovery time several hours). The parent species of the 13-cis like cycle absorb at 480 nm. That pigment fraction in the pellets whose absorption was not blue-shifted, also exhibited similar photoreactions to BR in suspension, but with an overall turnover rate only one order of magnitude slower. From a previous report (Lazarev and Terpugov, Biochim. Biophys. Acta 590 ,324–338,1980) and this one, it appears that the very slow photocycles in NaCl-BR of low moisture content originate from blue-shifted chromophores rather than from unchanged BR.     

ISOLATION OF PHOTOCHEMICAL REACTION CENTERS FROM A CAROTENOIDLESS MUTANT OF RHODOSPIRILLUM RUBRUM*

Lauryl dimethyl amine oxide was used to isolate reaction centers from a carotenoid-less mutant, strain G-9, of Rhodospirillum rubrum. These reaction centers have absorption spectra and light or chemically induced difference spectra very similar to those obtained from Rhodopseudomonas spheroides, strain R-26. But, unlike those from Rps. spheroides, they are more labile to higher detergent concentrations and to ammonium sulfate. The cytochrome content was estimated to be less than one per 10 P870.