ROSEOFLAVIN AS A BLUE LIGHT RECEPTOR ANALOG: SPECTROSCOPIC CHARACTERIZATION

Abstract— The blue light absorption band of roseoflavin is polarized along the axis roughly connecting N₃-C₈ positions. A weak, second π→π* transition with a polarization angle of ca. 25° is hidden under the short wavelength side of the blue absorption band. The excited state of roseoflavin is somewhat more basic than the ground state, by a 1.5 p K _a unit. The fluorescence quantum yield and lifetime of roseoflavin are substantially lower than those of other flavins, thus making it kinetically less efficient as a blue light photoreceptor.     

PHOTOSENSORY TRANSDUCTION IN CILIATES. II. POSSIBLE ROLE OF G-PROTEIN AND cGMP IN Stentor coeruleus

Abstract— The heterotrichous ciliate, Stentor coerulus , exhibits a welll defines photophobic response to a sudden increase in the intensity of visible light. the phobic reactions usually appear with a latency perios (i.e. a time delay between the onset of the stimulus and the stop response). This latency of phobic response was significatly increased when the cells werw incubated with 8-bromo-guanosine3',5'-cyclic monophospjhate. In the presence of this nucleotide, a reduction of cell responsiveness (i.e. the number of photophobically responding cells) was also observed. similar effects were observed when cells were treated with pertussis toxin, a G-protein activity modulator, and 3'-isobutyl-methylxanthine, an inhibitor of guanosine 3', 5'-cyclic monophosphate (cGMP) phosphodiesterase. the G-protein activator fluoroaluminate and 6-anilino-5,8-quinolinedione (LY 83583) (an effective agent for lowerin cellular cGMP levels) showed opposite effects on hte cell photophobic response. These result indirectly suggesnt that the level of cytoplamic cGMP, possibly modulated by a G-protein-coupled CGMP phosphodiesterase, plays a phototreasducing role in Stentor . In addition, using an antiserum raised against bovine transducin, a cross reacting protein with an apparent molecular mass of 39 kDa was detected on immunoblots. The α-subunits of a Stentor G-protein has also been partially cloned and sequenced. However, the possible coupling between the G-protein and the putative phosphodiesterase remains to be established.     

EFFECTS OF IONOPHORES AND RUTHENIUM RED ON THE PHOTOTAXIS OF STENTOR COERULEUS AS MEASURED BY SIMPLE DEVICES

Abstract— Two simple methods of phototaxis measurements have been applied to study the effects of ionophores on the negative phototactic response in Stentor coeruleus. The inhibitory effects of Ca²⁺-ionophore (A23187), Ca²⁺-blocking agent (Ruthenium Red), and K⁺ -ionophore (valinomycin) on photo-taxis have been determined. Results suggest that the influx of Ca²⁺ plays a transducing role in the phototaxis of Stentor.     

PHOTOSENSORY TRANSDUCTION IN CILIATES. IV. MODULATION OF THE PHOTOMOVEMENT RESPONSE OF Blepharisma japonicum BY cGMP

Abstract— The effect of various modulators of cytoplasmic guanosine 3',5'-cyclic monophosphate (cGMP) level on the step-up photophobic responses in Blepharisma japonicum has been investigated to clarify the possible role of cGMP in the mechanism of photosensory signal transduction. Membrane-permeable analogs of cGMP, 8-bromo-guanosine 3',5'-cyclic monophosphate or dibutyryl cGMP, caused a marked dose-dependent prolongation of the latency for the photophobic response, resulting in inhibition of the photophobic response in Blepharisma japonicum. A similar effect was observed when cells were treated with 3'-isobutylmethylxanthine (IBMX), a phosphodiesterase inhibitor, and pertussis toxin, a G-protein activity modulator. The G-protein activator, fluoroaluminate, and 6-anilino-5,8-quinolinedione (LY 83583), an agent which effectively lowers the cytoplasmic cGMP level, significantly enhanced the photoresponsiveness of these ciliates to visible light stimuli. These results suggest that cellular cGMP serves as a signal modulator in the photophobic response of Blepharisma japonicum.     

PROTECTION OF CHLOROPHYLL a BY CAROTENOID FROM PHOTODYNAMIC DECOMPOSITION

Abstract— The order of inhibition of the photooxidation of chlorophyll a in ethanol and ethanol-benzene is as follows: β-carotene, α-tocopherol, benzoquinone, DABCO, menadione, cholesterol and KI. The quenching of singlet oxygen by β-carotene occurs by a collisional quenching mechanism with a diffusion-controlled rate of 1.7 × 10¹⁰ M ^-1 s^-1. Photodecomposition of Chi a is faster in ethanol-D₂O than in ethanol-H₂O. Photoirradiation (660 nm) of the peridinin-Chl a -protein complex, a photosynthetic light-harvesting pigment isolated from marine dinoflagellates, did not show any photo-decomposition of its Chi a in H₂O or D₂O, even after an extended period (12 h) of irradiation. However, the carotenoid, peridinin, in the photosynthetic antenna pigment was photobleached (ca. 10%) during the irradiation. We conclude that the singlet oxygen formed as a result of the Chi photosensitization is immediately quenched by the low-lying triplet state of four peridinin molecules (per Chl a ) bound within the same protein crevice. The carotenoid thus effectively protects Chl a from photodynamic damage, providing a direct proof for the protective role of carotenoids in the photosynthetic pigment complex.     

ISOLATION AND CHARACTERIZATION OF THE PRESUMED PHOTORECEPTOR PROTEIN OF Blepharisma japonicum

Abstract— Photosensory responses in the ciliated protozoan Blepharisma japonicum are mediated by a hypericin-like chromophore, blepharismin, localized in granules distributed under the cell membrane. A blepharismin-binding protein, with an apparcnt molecular weight ranging between 35 and 38 kDa, has been isolated by means of column separations and preparative isoelectric focusing and characterized by means of gel electrophoresis, analytic isoelectric focusing as well as absorption and fluorescence spectroscopy.     

THE DISTANCE BETWEEN THE PHYTOCHROME CHROMOPHORE AND THE N-TERMINAL CHAIN DECREASES DURING PHOTOTRANSFORMATION. A NOVEL FLUORESCENCE ENERGY TRANSFER METHOD USING LABELED ANTIBODY FRAGMENTS

A novel antibody-fluorescence method has been developed to elucidate the chromophore topography in phytochrome as it undergoes a photochromic transformation. Förster energy transfer from N-terminal bound, fluorescently labeled Oat-25 Fab antibody fragments to the phytochrome chromophore was measured. The results suggest that the chromophore moves relative to the N-terminus upon the Pr → Pfr phototransformation. This conclusion is consistent with previous models which have proposed a reorientation and an interaction of the Pfr chromophore with the N-terminus. The method described appears to be the first study of a Forster energy transfer measurement using a donor-label attached to a Fab fragment of a photosensor protein.     

Electronic structures and spectra of halogenated purines and pyrimidines III.σ-Electronic structure and some comments on the mechanism of the xanthine oxidase reaction of 2-halopurines and related analogs

The extended HMO (EHMO) and Pople-Segal SCF-MO-CNDO /2 calculations on purine, 8-oxopurine, 2-oxopurine, 2-fluoropurine, and 2-chloropurine indicate significant polarizations of the σ cores. It is shown that the polarizations of both σ and π frameworks are mutually opposing in some cases. The results from the two methods are compared for these complex biomolecules. It is found that the EHMO calculations tend to over-polarize the σ and π frameworks. However, the CNDO shows anomalous π-electron densities on N₍₇₎ and C₍₈₎ of the purine rings, and the reason for this anomaly is not certain. The application of the results to the xanthine oxidase system indicates that the substrate molecules are subject to a specific orientation on the enzyme surface to counteract the electronic reactivity, in support of the previous prediction based on the π-electron calculations. The CNDO results appear to be more satisfactory than the EHMO in this respect.