ON THE MECHANISM OF WAVELENGTH REGULATION IN VISUAL PIGMENTS

Abstract— The contributions of different factors that might be responsible for the 500 nm absorption maximum of bovine rhodopsin are evaluated in detail. These include: (1) electrostatic interactions between the chromophore and a charged amino acid on the apoprotein; (2) exciton interactions with aromatic amino acids; (3) twisting about single bonds which have considerable double bond character; (4) weak interactions between the Schiff base and a putative counter-ion. Analysis of these mechanisms in terms of theoretical and experimental results suggests that(2–4) are all capable of contributing to the protein induced spectral shifts. However, the "external point charge" model proposed previously, i.e. mechanism (1), appears to provide the crucial interaction. In this paper, the theoretical basis for this model is discussed in detail. The model is briefly evaluated in light of the amino-acid sequence of bovine rhodopsin and possible implications for other visual pigments are considered.     

EFFECTS OF EXTRACELLULAR Ca2+ CONCENTRATION AND PAPAVERINE ON THE VISUAL CELL FUNCTION IN THE ISOLATED FROG RETINA

Abstract— Effects of extracellular Ca²⁺ concentration and papaverine on the PIII response of the electroretinogram and the dark adaptation process of the visual cells were studied in the isolated, aspartate-treated bullfrog retina. The amplitudes of both the fast and slow PIII responses are increased in 0.01 m M Ca²⁺ solution, but decreased in Ca²⁺-free solution containing 1 m M EDTA. The application of 0.1 m M papaverine in the presence of 1 m M Ca²⁺ led to the enhancement of the slow PIII response at lower stimulus intensity and the prolongation of the slow PIII response, but these effects of papaverine on the response were lost when Ca²⁺ was removed from the bathing fluid. The half-time of recovery of the fast PIII response amplitude after switching off the adapting light was a linear function of the amount of bleached rhodopsin. Papaverine in the absence of Ca²⁺ produced about 2-fold increase in the half-time of recovery of the response. These findings suggest that chemical reactions which are sensitive to papaverine in the absence of Ca²⁺ are implicated in the dark adaptation process of the visual cells.     

CHANGE IN THE BASE COMPOSITION OF MESSENGER RNA OF ESCHERICHIA COLI BY ULTRAVIOLET IRRADIATION AND ITS RECOVERY

Abstract— The base composition of messenger RNA in Escherichia coli B/r and B _8–1 irradiated with ultraviolet (u.v.) light has been examined. The experimental results are as follows: (1) the synthesis of rapidly labeled RNA does not stop in ultraviolet irradiated bacteria. (2) The rapidly labeled RNA in irradiated cells shows a change in base composition corresponding to the formation of pyrimidine dimers in DNA molecules. The mole per cent of adenine component is increased with ultraviolet dose. The ratio of purine/pyrimidine becomes larger and the GC content smaller. (3) The base composition of the rapidly labeled RNA in irradiated bacteria reversed to that in unirradiated cells, when the irradiated cells were reactivated by experimental procedures for photoreactivation or dark reactivation. The reversion in the base composition corresponds well to the decrease in the amount of thymine dimers in DNA molecules. (4) The mechanism of the change in the base composition of rapidly labeled RNA caused by ultraviolet irradiation is discussed.     

超声辐照前后水煤浆浆体的动电势变化研究

选用大同煤和神木煤制备水煤浆，考察了超声辐照前后水煤浆浆体的动电势变化。实验结果表明，煤浆经超声辐照后添加剂在煤粒表面的饱和吸附量增加，相应煤浆浆体的动电势也随之增加，而添加剂在煤粒表面饱和吸附量的增加与超声辐照引起的煤粒比表面积的增加密度相关。动电势增加的另一原因与辐照后煤浆的电导率增加有关。超声辐照导致煤浆浆体静态稳定性大幅度提高的部分原因是由于动电势的增加。     

THYMINE DIMERIZATION IN L-STRAIN MAMMALIAN CELLS AFTER IRRADIATION WITH ULTRAVIOLET LIGHT AND THE SEARCH FOR REPAIR MECHANISMS

Abstract— The formation of thymine dimers in the DNA of L -strain mammalian cells after irradiation with ultraviolet light has been demonstrated. The amount of dimer formed rises with the dose of u.v. light.
In the course of post-irradiation incubation the thymine dimers remain in the TCA insoluble fraction and diminish as did the other thymidine-H³ derivatives with increasing incubation time. The dimer is not found in the soluble fraction. Thus, dimer excision (i.e. its liberation into the soluble fraction) as an expression of repair of radiation damage analogous to dark repair in E. coli was not found in these experiments.     

ON THE POSSIBILITY OF PROTEIN–CHROMOPHORE CHARGE TRANSFER IN VISUAL PIGMENTS

Abstract— Quantum chemical calculations have been performed on the retinylidene Schiff base assumed to be protonated or hydrogen bonded at the nitrogen and perturbed by a carboxylate ion at its β-ionone end. The results indicate that weak charge transfer type bands might exist due to electron transfer from the carboxylate ion to the Schiff base and that the intense (π, π*) band in the visible spectrum might acquire a significant degree of charge transfer character.     

CO-OPERATION OF PERIPHERAL AND INTEGRAL MEMBRANE PROTEINS IN THE LIGHT DEPENDENT ACTIVATION OF ROD GTPase AND PHOSPHODIESTERASE

Abstract— During the purification of light activated GTPase, we have observed that the GTPase activity is suddenly lost because of the separation of two components which are both needed for enzymatic activity. The G fraction or protein (apparent mol. wt 55000) can selectively bind GTP and is necessary for PDE activation by illuminated rhodopsin. However, the G fraction shows no GTPase activity unless it is supplemented by illuminated rhodopsin and the H fraction, a heat labile, trypsin resistant macro-molecule with an apparent mol. wt of 53 000. The G and H fractions (and GTPase activity) show a significant increase in binding to bleached as compared with unbleached disc membranes.     

CHROMOPHORE OF A LONG-LIVED PHOTOPRODUCT FORMED WITH METARHODOPSIN III IN THE ISOLATED FROG RETINA

Abstract Long-lived photoproducts of frog rhodopsin in isolated retina and digitonin solution have been investigated by spectrophotometry and their chromophores have been analyzed by high-pressure liquid chromatography (HPLC). By irradiation (> 560 nm) at 3°C and pH 8.6, a product analogous to metarhodopsin III (MIII) is formed, whose absorption maximum is at about 450 nm. This product decays more slowly than MIII does. The results of HPLC analysis indicate that the chromophore of this photoproduct is 7- cis retinal and that of MIII is all-trans retinal. The product possessing 7- cis retinal is called 7- cis photoproduct. The amount of 7- cis isomer in rhodopsin solution irradiated at various temperatures between 15°C and –82°C, has been determined. The results suggest that the 7- cis photoproduct can be formed by the photoconversion of lumirhodopsin and metarhodopsin I.     

LIGHT-INDUCED INFRARED LIGHT SCATTERINGSIGNAL IN THE RETINA: EFFECT OF PHOSPHODIESTERASE INHIBITORS

Visible light changes IR light scatter through a toad retina. This signal presents three components: at low light intensity (100-400 bleached rhodopsins/rod) an early decrease in IR light scatter, of small amplitude, with time to peak of 1-6 s; at intermediate light intensity (1200-16,000 bleached rhodopsins/rod) a slow increase in IR light scatter, with time to peak of 10-30 s; at high light intensity (50,000-160,000 bleached rhodopsins/rod) a last increase in IR light scatter, with time to peak of 1 min. Light sensitivity, amplitude and time to peak of the last two components are increased by inhibitors (3-isobutyl-1-methyl-xanthine and papaverine) of the cyclic 3'5' guanosine monophosphate phosphodiesterase.     

EFFECTS OF RHODAMINE 123 IN THE DARK AND AFTER IRRADIATION ON MITOCHONDRIAL ENERGY METABOLISM

Isolated rat liver mitochondria have been used to study the mechanism of toxicity of Rhodamine 123 (Rho 123) in the dark and after irradiation with visible light. We report an inhibition of adenosine 5'-diphosphate phosphorylation which is increased after illumination. In the dark, the first steps of the phosphorylation process (i.e. the entry of substrates into the matrix, the electron transport to oxygen and the creation of the proton gradient) as well as ATPase activity are not significantly perturbed at Rho 123 concentration below 10 micrograms/mL. In contrast, the movements of the phosphate compounds are drastically impaired. Irradiation strengthens the detrimental effects in an oxygen dependent process. The nature of the noxious transient species is not clearly established, but it is suggested that singlet oxygen could be responsible for the observed damage.     

INFRARED AND RAMAN STUDIES ON SOME IMINES AND THEIR PICRATES. RELATION TO THE PROBLEM OF PROTONATION IN VISUAL PIGMENTS

Abstract— Picrates of conjugated Schiff bases have C=N stretching frequencies similar to those of the respective free bases. This raises the question if the frequency of this vibration can provide proof for the protonation of the retinylidene Schiff base occurring in visual pigments.     

LIGHT DAMAGE IN THE RAT RETINA: EFFECT OF A RADIOPROTECTIVE AGENT (WR-77913) ON ACUTE ROD OUTER SEGMENT DISK DISRUPTIONS

Primary events in the course of light induced retinal lesions are still not fully elucidated. Under chronic conditions, lipid peroxidation in the retina and death of photoreceptor cells are observed. The radioprotective agent WR-77913 scavenges singlet oxygen, hydrated electrons and free radicals. WR-77913 was used to protect against acute light induced photoreceptor outer segment membrane disruptions in the rat retina. There was a partial but not complete protection at higher illuminance levels (800 lx for 30 min), whereas threshold lesions (400 lx for 30 min) were almost completely prevented. These observations indicate an involvement of photodynamic reactions in causing acute photoreceptor lesions.     

PHOTOLYSIS AND PHOTO-OXIDATION OF AMINO ACIDS AND PEPTIDES—VI. A STUDY OF THE INITIATION OF DISULPHIDE INTERCHANGE BY LIGHT IRRADIATION

Abstract— Investigations concerning the occurrence of disulphide interchange for disulphides related to cystine are made with either a single unsymmetrical disulphide or an equimolar mixture of two symmetrical disulphides. It is found that in the pH range 1–6 where spontaneous interchange can usually be disregarded, interchange can be initiated by the addition of a small amount of a thiol or by irradiation with ultraviolet light from either of two artificial sources, interchange being more rapid at pH 4–6 than at pH 1–3. Direct sunlight and, to a lesser extent, visible (filtered tungsten) light are also effective. Disulphide interchange in the solid state can also be promoted by ultraviolet light. A comparison is made between the roles' of S-S and C-S bond fission in the photochemistry of disulphides, with respect not only to the promotion of disulphide interchange, but to other phenomena such as the dismutation of disulphides to thioethers and trisulphides.     

VIABILITY OF THE SPORES FORMED AFTER UV IRRADIATION IN DICTYOSTELIUM DISCOIDEUM

Both abilities of germination of spores formed after UV irradiation and of growth of amoeboid cells emerged from the spores were studied on two kinds of Dictyostelium discoideum strains, NC-4 and ys-13.
An inhibition of germination was observed on the spores of ys -13 when formed after UV irradiation, while no inhibition was detected on the ability of germination of spores of NC-4. The amoeboid cells of ys -13 emerged from the spores showed a heavy delay of growth, although no delay of growth was detected even on the amoeboid cells of NC-4 emerged from the spores formed after UV irradiation. The strain of NC-4 must repair UV lesions fully before spore formation, while the spores of ys-13 must keep some UV lesions unrepaired and send them to the next generation of amoeboid cells. The characters of UV lesion inheritable through the spores to the next amoeboid cells in ys-13 were discussed.     

ON THE STATE OF CHROMOPHORE PROTONATION IN RHODOPSIN: IMPLICATION FOR PRIMARY PHOTOCHEMISTRY IN VISUAL PIGMENTS

Resonance Raman multicomponent spectra of bovine rhodopsin, isorhodopsin, and bathorhodopsin are obtained at low temperature. Application of the double beam, 'pump-probe' technique allows an extraction of the rhodopsin and bathorhodopsin spectra in both protonated and deuterated media. Our results show that the Schiff bases of both rhodopsin and bathorhodopsin are fully protonated and the degree of protonation is unaffected by the rhodopsin-bathorhodopsin transformation. Further, the data support the concept or cis-trans isomerization as occurring in this transition. The effect of these results on various models for the primary photochemical event in vision is discussed.     

OXIDATION OF RIBOSE MOIETY OF ADENOSINE INDUCED BY IRRADIATION OF NEAR-UV LIGHT IN THE PRESENCE OF IRON(III) CHLORIDE

Abstract— Irradiation of near-UV light in the presence of FeCl₃ caused oxidation of adenosine with concomitant reduction of Fe(III) under either aerobic or anaerobic atmosphere. Major photoproducts were adenine and adenosine-5'-aldehyde, showing that oxidation took place mostly at the ribose moiety of adenosine. Detailed study showed that the reaction proceeded by the light absorption due to Fe(III) at 300-350 nm.     

CAROTENOID PIGMENTS AND THE IN VIVO SPECTRUM OF BACTERIOCHLOROPHYLL

Abstract— The isolation of a mutant, strain PM-9, of Rhodopseudonionus spheroides with an abnormal complement of carotenoid pigments is described.
PM-9 accumulates phytoene, phytofluene, ζ-carotene and neurosporene. Semi-aerobic cultures form more ζ-carotene and neurosporene relative to total carotenoids than do photo-synthetic cultures.
PM-9 is killed on exposure to light and oxygen.
By making use of the effect of oxygen on the nature of the carotenoids in PM-9, we have shown that these pigments do not directly influence the in vivo spectrum of bacteriochlorophyll.
Diphenylamine inhibits the synthesis of coloured carotenoids in Rhodopseudonionos gelatinosa but does not change the bacteriochlorophyll spectrum.     

二氯二茂钛引发二苯乙炔聚合的研究

在高温下 ,二氯二茂钛受光照引发二苯乙炔聚合 ,得到结晶度为 95 %的聚二苯乙炔 .经红外、质谱、热分析、X光衍射、元素分析等方法 ,确定其结构为顺顺式螺旋刚性棒状结构 ,形成假六方晶系堆积 ,螺旋柱直径即螺旋轴间距为 1 46nm ,其特征红外吸收峰位于 1179cm-1和 115 6cm-1.在聚合引发阶段中止反应 ,分离出聚合中间体六苯基苯 .根据不同温度时聚合转化率与时间的关系曲线 ,测得聚合活化能为 2 6× 10 5J/mol.热分析表明 ,结晶聚二苯乙炔在 493K至 5 73K比较稳定 ;在 713K的吸热峰显示出破坏晶格的能量 .根据聚合反应诱导期长、中间体和聚合物的结构以及阻聚反应 ,推断聚合过程中二氯二茂钛受光照产生二氯化钛 ,催化二苯乙炔进行均相定向聚合 .结晶聚二苯乙炔电导率小于 10 -12 (Ωcm) -1.     

SINGLE-STRAND BREAKS IN THE DNA OF THE uvrA AND uvrB STRAINS OF ESCHERICHIA COLI K-12 AFTER ULTRAVIOLET IRRADIATION

Abstract— DNA single-strand breaks were produced in uvrA and uvrB strains of E. coli K-12 after UV (254 nm) irradiation. These breaks appear to be produced both directly by photochemical events, and by a temperature-dependent process. Cyclobutane-type pyrimidine dimers are probably not the photoproducts that lead to the temperature-dependent breaks, since photoreactivation had no detectable effect on the final yield of breaks. The DNA strand breaks appear to be repairable by a process that requires DNA polymerase I and polynucleotide ligase, but not the recA, recB, recF, lexA 101 or uvrD gene products. We hypothesize that these temperature-dependent breaks occur either as a result of breakdown of a thermolabile photoproduct, or as the initial endonucleolytic event of a uvrA , uvrB -independent excision repair process that acts on a UV photoproduct other than the cyclobutane-type pyrimidine dimer.     

THE MODIFICATION OF RIBONUCLEASE-A BY NEARULTRAVIOLET IRRADIATION IN THE PRESENCE OF PSORALEN

Abstract— Ribonuclease A is inactivated when irradiated under oxygen by UV-A light in the presence of psoralen. The rate of inactivation is greatly reduced by sodium azide. ascorbate or nitrogen, whereas the substrate gives only very limited protection. A ribonuclease sample modified to 40% remaining activity presented a significant modification of amino acid residues known to be sensitive to oxidation and 1.4 mol of bound psoralen per mol of protein. The secondary structure of the enzyme, as assessed by circular dicroism was not changed by irradiation; neither was aggregation of the enzyme to a higher mol wt evident. Studies on the tryptic peptides fractionated by high performance liquid chromatography showed that the photomodification occurs with very low selectivity. All the five peptides containing hystidine, tyrosine and methionine residues were greatly modified, although two, those containing histidine residues 12 and 119 in the sequence, amino acids known to be involved in the catalytic activity of ribonuclease. are modified to a greater extent. The protein bound psoralen. revealed by radioactivity in the HPLC eluate, was not found associated to only one or few peptide peaks but spread on a large zone of elution.