Die kurzzeitigen ultravioletten Differenzspektren bei der Photosynthese

1968 ◽  
Vol 23 (2) ◽  
pp. 220-224 ◽  
Author(s):  
H. H. Stiehl ◽  
H. T. Witt

Plastoquinone PQ is engaged in photosynthesis 1. Difference spectra in the UV-region indicate that PQ is an intermediate in the electron transport chain 2. PQ is located as a pool between the two light reactions I and II 3. PQ is reduced by hνII and oxidized by hνI3. In this paper the difference spectra which occur during light excitation of spinach chloroplasts and chlorella vulgaris were measured in the UV-region with high resolution by the high sensitive method of periodical flash photometry 6.On excitation with long flashes (10—1 sec) the difference spectra are similar to those obtained when plastoquinone is reduced to hydroquinone in vitro (see figs. 1, 3 and 6). Deviations in the case of chlorella (fig. 4) are caused by additional NADP-reduction. After extraction of plastoquinone from chloroplasts the difference spectra do not occur during light excitation but they can be produced in full after reconstitution with synthetic plastoquinone A (see fig. 2).In the presents of far red background light (718 nm) which excites only light reaction I the magnitude of the spectrum is doubled (see fig. 3).By excitation with short flashes (10—5 sec), two different spectra were found. The difference spectrum with a life-time of 5 · 10—4 sec (fig. 7) is new and does not correspond to that of plastoquinone in vitro. The difference spectrum with a life-time of about 2·10—2 sec (fig. 5) corresponds to the plastoquinone reduction. The magnitude of this spectrum is ten times smaller than that obtained by excitation with long flashes (fig. 6).The 1:10 ratio of the magnitude of the spectra in short and long flashes can be interpreted by a pool of plastoquinone between the two light reactions with a dynamic capacity of ten electrons. The doubled magnitude of the spectra in far red background light can be interpreted by an electron acceptor pool for plastoquinone with a capacity of five electrons (see also the following papers).

1964 ◽  
Vol 19 (8) ◽  
pp. 693-707 ◽  
Author(s):  
B. Rumberg ◽  
H. T. Witt

The primary reactions of photosynthesis are fast reactions. To get detailed informations we developed three different methods. 1. sensitive flash photometry 38, 2. periodically chemical relaxation51, 26a, 3. manometry in flashing light groups 26b. With the method of flash photometry fast absorption changes in suspensions of chlorella and spinach chloroplasts were studied. 7 different types of absorption changes have been separated and analysed (Fig. 2). Older results 28-49lead to a reaction scheme published in l. c. 13. This scheme was refined (s. Fig. 3) by results published in l. c. 14-26. In the following 6 papers these investigations are described in detail and supplement by new results.Separation of the difference-spectrum of chlorophyll-al (P 700) in 5 ways. Under “normal” conditions (exciting Hill - active chloroplasts with blue light between 380 and 480 mμ or with red light between 620 and 720 mµ) mixed changes of absorption can be observed between 400 and 800 mµ (Fig. 4, this difference-spectrum does not include the changes with τ<10-3 sec, s. Fig. 2). Out of this overall difference-spectrum one component with changes of absorption at 430 and 703 mμ could be separated by using the following different systems: a) aged chloroplasts reactivated by addition of reduced DPIP or reduced PMS (ascorbate in excess) (Fig. 5), b) plastoquinone-extracted chloroplasts [extraction with petroleumether] (Fig. 6), c) digitonin-treated chloroplasts reactivated by addition of reduced PMS [ascorbate in excess] (Fig. 7), d) chloroplasts at -150°C with addition of reduced PMS [ascorbate in excess] (Fig. 8), e) chloroplasts under the influence of far-red background-light [728 mµ] (Fig. 9).Kinetics. During the flash the absorption at 430 and 703 mµ decreases very fast (<10-5 sec). In the dark the back reaction takes place in ≈10-2 sec at 20°C (s. Fig. 10). This reaction time is always observable in the presents of far redbackground light λ>700 mμ. (The detecting light at 703 mµ can act already as far red background light.) At very low itensities of far red background light the backreaction takes place however in <10-4 sec (details s. l. c. 25a).Identification of chlorophyll-al. The upper results (5 equal spectra under different conditions) suggest that the changes of absorption at 430 and 703 mμ are caused by one substance. This was additionally proved by comparing the magnitude and the kinetic of both changes in reactivated aged or digitonin-treated chloroplasts under different conditions. The ratio of the amplitudes and the halflifes are identical at 430 and 703 mµ at different values of pH (Fig. 11 and Fig. 12) and also at different concentrations of added reduced PMS (Fig. 13). Decreases of absorption just within the two absorption bands of chlorophyll-a indicate that very probably a chlorophyll-a (Chl-aI-430-703) is in action. From the magnitude of the changes of absorption at 703 mμ it follows, that Chl-al has a concentration of 0,1% of the bulk of chlorophyll.Oxidation of Chl-ai. The decreases of absorption indicate an oxidation of Chl-al in the light. This is proved by the fact, that in aged or digitonin-treated chloroplasts reduced PMS can be directly coupled to the light product. This is demonstrated by the strong acceleration of the back reaction with increasing concentrations of reduced PMS (Fig. 13 and Fig. 15) and by the demonstration of a first order back reaction (Fig. 14 and Fig. 15). The experimental results are theoretically explained. It is proved, that the light product Chl-aI⨁ reacts with PMSH⊖ [compare measuring points and theoretical curve in Fig. 12] [s. scheme (1)]. From the measurements it follows a reaction constant of 1,5·107 l/Mol·sec (Fig. 15) and an energy of activation of 3,8 kcal/Mol (Fig. 16) for the reaction of Chl-aI⨁ with PMSH⊖.Chl-aI-oxidation as a primary act. The fact, that Chl-ai⊕ is built up within <10-5 sec (20°C) [Fig. 10] and that Chl-aI⨂ could be trapped at -150°C (Fig. 8 and Fig. 17) give evidence, that this oxidation is a primary act. The electron acceptor of Chl-ai is called Ζ. Ζ⊖ reduces via intermediary products TPN. Under the reported conditions a - e one light reaction cycle (I) of the overall electron transport system of photosynthesis has been isolated [s. scheme (1) and (2)].


1973 ◽  
Vol 19 (10) ◽  
pp. 1235-1238
Author(s):  
Andrew M. B. Kropinski ◽  
Joyce Boon ◽  
Rozanne Poulson ◽  
W. James Polglase

A pigment absorbing at 503 nm (P503) was observed in the difference spectra of several strains of microorganisms. The pigment was present in most facultative anaerobes and absent from many but not all of the aerobes examined. P503 is probably not a component of the normal oxygen-linked electron transport chain since the pigment was present in both respiratory sufficient (ρ+) and deficient (ρ−) strains of Saccharomyces cerevisiae. There was no apparent correlation between P503 and either the cytochromes or any known metabolic cellular activities. However, the widespread occurrence of P503 in microorganisms suggests that it is of functional significance.


1980 ◽  
Vol 35 (1-2) ◽  
pp. 139-144 ◽  
Author(s):  
Marc Symons ◽  
Anthony Crofts

Abstract The flash-induced carotenoid bandshifts have been studied for various strains of both Rho-dopseudomonas sphaeroides and capsulata. A technique for calculating shifts of isobestic points down to 0.05 nm is described. To this end, special attention has been paid to the appropriate correction for reaction center absorbance changes occuring concommitantly with the carotenoid bandshifts. Plots of the wavelength of the isobestic point versus the corresponding absorption changes at the maximum of the difference spectrum have been made, suggesting the existence of different pools of carotenoids. The various pools of carotenoids seem to have different sizes, inducing non-linearities in the plots. In some cases spectral differences of the pools have to be assumed. A possible interpretation of the results would be that each electrogenic span of the electron transport chain is to be associated with its own pool of carotenoids, all the pools behaving in somewhat independant way. We discuss possible difficulties in making those measurements.


1959 ◽  
Vol 234 (8) ◽  
pp. 2176-2186 ◽  
Author(s):  
F. Edmund Hunter ◽  
Jerome F. Levy ◽  
Joan Fink ◽  
Beverly Schutz ◽  
Francisco Guerra ◽  
...  

1970 ◽  
Vol 37 (2) ◽  
pp. 259-267 ◽  
Author(s):  
G. C. Cheeseman ◽  
Dorothy J. Knight

SummaryThe dissociation of casein aggregates by the detergent sodium dodecyl sulphate (SDS) gave rise to difference spectra and these spectra were characteristic for each of the different types of casein. Increase in absorption by the chromophore groups, tyrosine and tryptophan, when αs1- and β-casein aggregates were dissociated indicated binding of the detergent at regions of the molecule containing these residues. A decrease in absorption when κ-casein was dissociated indicated that the tyrosine and tryptophan residues were not in the region of the molecule to which the detergent was bound and that in the κ-casein aggregate these residues were in a more hydrophobic environment. Peaks on the difference spectra were obtained at 280 and 288 nm for αs1-casein and 284 and 291 nm for β-casein and troughs at 278 and 286 nm for κ-casein. The difference spectrum reached a maximum value when the αsl- and β-casein aggregates were dissociated and the further binding of SDS did not alter this value. The large negative change in the difference spectrum of κ-casein did not occur until after most of the aggregates were dissociated and did not reach a maximum until binding with SDS was complete. The value obtained for ΔOD was found to be temperature-dependent for β-casein-SDS interaction, but not for αs1- and κ-casein. Changes in spectra were also observed when αs1- and κ-casein interacted to form aggregates. The data obtained confirmed the importance of hydrophobic binding in casein aggregate formation and indicated the possible involvement of tyrosine and tryptophan residues in this binding.


1972 ◽  
Vol 129 (3) ◽  
pp. 755-761 ◽  
Author(s):  
M. V. Jones ◽  
D. E. Hughes

In cell-free extracts of Pseudomonas ovalis nicotinic acid oxidase is confined to the wallmembrane fraction. It is associated with an electron-transport chain comprising b- and c-type cytochromes only, differing proportions of which are reduced by nicotinate and NADH. CO difference-spectra show two CO-binding pigments, cytochrome o (absorption maximum at 417nm) and another component absorbing maximally at 425nm. Cytochrome o is not reduced by NADH or by succinate but is by nicotinate, which can also reduce the ‘425’ CO-binding pigment. The effects of inhibitors of terminal oxidation support the idea of two terminal oxidases and a scheme involving the ‘425’ CO-binding pigment and the other components of the electron-transport chain is proposed.


Author(s):  
RAFAELA M. ALVARIZ ◽  
ISABEL T.D.S. MOREIRA ◽  
GABRIELA K. CURY ◽  
CARMEN R. VARGAS ◽  
ALETHÉA G. BARSCHAK

2020 ◽  
Vol 319 (1) ◽  
pp. R69-R78
Author(s):  
Zhuping Jin ◽  
Quanxi Zhang ◽  
Eden Wondimu ◽  
Richa Verma ◽  
Ming Fu ◽  
...  

The production of H2S and its effect on bioenergetics in mammalian cells may be evolutionarily preserved. Erythrocytes of birds, but not those of mammals, have a nucleus and mitochondria. In the present study, we report the endogenous production of H2S in chicken erythrocytes, which was mainly catalyzed by 3-mercaptopyruvate sulfur transferase (MST). ATP content of erythrocytes was increased by MST-generated endogenous H2S under normoxic, but not hypoxic, conditions. NaHS, a H2S salt, increased ATP content under normoxic, but not hypoxic, conditions. ATP contents in the absence or presence of NaHS were eliminated by different inhibitors for mitochondrial electron transport chain in chicken erythrocytes. Succinate and glutamine, but not glucose, increased ATP content. NaHS treatment similarly increased ATP content in the presence of glucose, glutamine, or succinate, respectively. Furthermore, the expression and activity of sulfide:quinone oxidoreductase were enhanced by NaHS. The structural integrity of chicken erythrocytes was largely maintained during 2-wk NaHS treatment in vitro, whereas most of the erythrocytes without NaHS treatment were lysed. In conclusion, H2S may regulate cellular bioenergetics as well as cell survival of chicken erythrocytes, in which the functionality of the electron transport chain is involved. H2S may have different regulatory roles and mechanisms in bioenergetics of mammalian and bird cells.


Author(s):  
J. Bruley ◽  
D. B. Williams

This paper concerns the influence of sample thickness on spatial-difference spectra, and seeks to identify if an interface dependent signal may be generated as an artifact of grain boundary grooving. The spatial-difference profiling technique may be used to identify variations in composition and electronic structure across interfaces at sub-nanometer length scales. The signal-to-background ratios and hence visibility of small changes to the near-edge structure and edge intensities are enhanced using this technique by removing intense energy dependent backgrounds. These backgrounds are assumed to be only slowly varying with respect to the electron probe position. A spatial-difference spectrum is generated from the difference between two spectra after suitable normalization or scaling. This scaling is achieved by either matching intensities of the background prior to a characteristic absorption edge (for compositional profiles) or by normalizing to some characteristic structure of the near-edge structure (for bonding profiles). The latter is performed typically after subtraction of a smooth power-law background modeled in the region immediately preceding the edge.


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