New chlorophyll-b forms in a chlorophyll-detergent photosynthetic model system

The absorption spectra of chlorophyll b in Triton X-100 micelles at room temperature are superpositions of components with absorption maxima at 640.8, 648.9, 659.5, 669.6, 682.1 and 695.7 nm, obtained from Gaussian analysis of the spectra. The last four forms strongly overlap the chlorophyll a forms of this system obtained with maxima at 659.3, 667.6, 674.3, 680.8, 686.5, 692.8, 701.9, 713.6 and 722.0 nm.

Since the in vivo chlorophyll a forms practically coincide with the forms found in this system, the possible existence of in vivo overlapping chlorophyll b and a forms eventually should be taken into consideration. In this case, however, the Gaussian analysis of in vivo absorption bands in itself in the proper spectrum range cannot discriminate between chlorophyll a and b components.     

A study of chlorophyll b in vivo

1. The absorption spectrum of chlorophyll b in vivo at 77°K is presented as the difference spectrum between preparations of spinach and chlorophyll b-free Vischeria stellata chloroplasts.

2. A shoulder on this spectrum around 662 nm is due to a component different from chlorophyll b. This component may well be identical with the chlorophyll a form, chlorophyll a (665).

3. The 77°K chlorophyll b absorption spectra in the nonfractionated photosyn-thetic pigment apparatus and in fractions mainly representing Photosystems 1 or 2 are not significantly different.

4. The aerobic irreversible photobleaching of chlorophyll b was studied in the intact pigment complex as well as in fractions mainly consisting of Photosystem 1 or 2. A two-step photobleaching was observed in all cases. The time-course of this bleaching was not significantly different for chlorophyll b in both fractions.

5. These results do not indicate that more than a single chlorophyll b complex occurs in vivo.     

Intergeneric structural variability of the primary donor of photosynthetic bacteria: Resonance raman spectroscopy of reaction centers from two Rhodospirillum and Rhodobacter species

Several structural properties of the primary donor of Rhodobacter sphaeroides, wild type, have been obtained recently from difference resonance Raman spectra (Robert, B. and Lutz, M. (1986) Biochemistry 5, 2303–2309). In order to test the interspecific variability of the primary donor structure, we extended those observations to reaction centers of Rhodospirillum rubrum, wild type, using the same difference methods. Difference resonance Raman spectra of the primary donor are extremely close to those obtained for Rhb. sphaeroides. Identical conclusions can be drawn for the two species about the molecular interaction states of the magnesium atoms and of the acetyl carbonyl groups of both bacteriochlorophyll a molecules constituting their primary donors. However, the keto carbonyl of one of the bacteriochlorophylls, which is intermolecularly bound in Rhb. sphaeroides, is free from bonding in Rsp. rubrum. It thus appears that there is a limited interspecific variability of the structure of the primary donor in Rhodospirillales. These results also demonstrate an interspecifically variable, environmental asymmetry around the two primary donor molecules, which should variably affect charge distributions over these two molecules, in transient intermediates states of P, according to the species considered.     

Excitation energy transfer in the light-harvesting chlorophyll

The “light-harvesting chlorophyll a/b · protein” described by Thornber has been prepared electrophoretically from spinach chloroplasts. The optical properties relevant to energy transfer have been measured in the red region (i.e. 600–700 nm). Measurements of the absorption spectrum, fluorescence excitation spectrum and excitation dependence of the fluorescence emission spectrum of this protein confirm that energy transfer from chlorophyll b to chlorophyll a is highly efficient, as is the case in concentrated chlorophyll solutions and in vivo. The excitation dependence of the fluorescence polarization shows a minimum polarization of 1.9 % at 650 nm which is the absorption maximum of chlorophyll b in the protein and rises steadily to a maximum value of 13.8 % at 695 nm, the red edge of the chlorophyll a absorption band. Analysis of these measurements shows that at least two unresolved components must be responsible for the chlorophyll a absorption maximum. Comparison of polarization measurements with those observed in vivo shows that most of the depolarization observed in vivo can take place within a single protein. Circular dichroism measurements show a doublet structure in the chlorophyll b absorption band which suggests an exciton splitting not resolved in absorption. Analysis of these data yields information about the relative orientation of the S₀→S₁ transition moments of the chlorophyll molecules within the protein.     

Composition of the photosystems and chloroplast structure in extreme shade plants

Chloroplasts were isolated from leaves of three species of tropical rainforest plants, Alocasia macrorrhiza, Cordyline rubra and Lomandra longifolia; these species are representative of extreme “shade” plants. It was found that shade plant chloroplasts contained 4–5 times more chlorophyll than spinach chloroplasts. Their chlorophyll a/chlorophyll b ratio was 2.3 compared with 2.8 for spinach. Electron micrographs of leaf sections showed that the shade plant chloroplasts contained very large grana stacks. The total length of partitions relative to the total length of stroma lamellae was much higher in Alocasia than in spinach chloroplasts. Freeze-etching of isolated chloroplasts revealed both the small and large particles found in spinach chloroplasts.

Despite their increased chlorophyll content, low chlorophyll a/chlorophyll b ratio, and large grana, the shade plant chloroplasts were fragmented with digitonin to yield small fragments (D-144) highly enriched in Photosystem I, and large fragments (D-10) enriched in Photosystem II. The degree of fragmentation of the shade plant chloroplasts was remarkably similar to that of spinach chloroplasts, except that the subchloroplast fragments from the shade plants had lower chlorophyll a/chlorophyll b ratios than the corresponding fragments from spinach. The D-10 fragments from the shade plants had chlorophyll a/chlorophyll b ratios of 1.78-2.00 and the D-144 fragments ratios of 3.54–4.07. We conclude that Photosystems I and II of the shade plants have lower proportions of chlorophyll a to chlorophyll b than the corresponding photosystems of spinach. The lower chlorophyll a/chlorophyll b ratio of shade plant chloroplasts is not due to a significant increase in the ratio of Photosystem II to Photosystem I in these chloroplasts.

The extent of grana formation in higher plant chloroplasts appears to be related to the total chlorophyll content of the chloroplast. Grana formation may simply be an means of achieving a higher density of light-harvesting assemblies and hence a more efficient collection of light quanta.     

Preparation and properties of the water-soluble chlorophyll-bovine serum albumin complexes

By mixing chlorophyll (Chl) a or b with a dense bovine serum albumin solution, the water-soluble Chl-bovine serum albumin complexes were prepared. These complexes, eluted near the void volume on a gel filtration, were separated well from unreacted bovine serum albumin, indicating an aggregation of such molecules in the complexes. Preparation of chlorophyllide (Chlide) a- or Chlide b-bovine serum albumin complex was unsuccessful, while the phytol-, and β-carotene-bovine serum albumin complexes could be obtained. Chls in the Chl-bovine serum albumin complexes had the following characteristics. (i) Main absorption peak of Chl a or b in the red region occurred at 675 nm or 652 nm, respectively. The Chl a-bovine serum albumin complex having absorption peak at 740 nm was also prepared. As compared with the stabilities of Chl a and b in Triton X-100. (ii) Both Chls in the bovine serum albumin-complexes were stable against oxidative stresses, such as photobleaching, Fenton reagent, peroxidase-H₂O₂ system. But (iii) they were easily hydrolyzed by chlorophyllase. These properties of Chls in the bovine serum albumin-complexes were similar to those of Chls in the isolated light-harvesting Chl a/b protein complex. A possible localization of Chls within the bovine serum albumin complexes was suggested that the porphyrin moiety of Chl was buried in bovine serum albumin; however, the hydrophilic edge of porphyrin ring, adjacent to the phytol group, occurred in the hydrophilic region of a bovine serum albumin molecule.     

Protein-protein interactions of the light-harvesting chlorophyll a/b protein. II. Evidence for two stages of cation independent association

In a previous paper, we observed a two-stage cation-independent association of the light-harvesting chlorophyll a/b protein from spinach chloroplasts based on concentration-dependent changes in the sedimentation coefficient. The two stages of association occurred between (2–4) and (4–7) μg/ml chlorophyll. In this paper, we provide further evidence for this association.

This includes: (1) A decrease in the number of divalent cation binding sites in the second stage of association. (2) A corresponding decrease in the extent of the cation-dependent association. (3) A positive deviation from Beer's law for chlorophyll b for both stages of the cation-independent association and a positive deviation for chlorophyll a for the second stage of association only. (4) A change in the fluorescence emission of both chlorophyll a and b. The change for chlorophyll b was observed for both steps of association whereas that for chlorophyll a was observed for the second step of association only. Therefore, the first stage of association affects only chlorophyll b whereas the second stage alters the environment of both chlorophyll a and b. (5) In addition, divalent cations quenched chlorophyll fluorescence. However, the quenching which required 200–300 μM divalent cation for half-maximal effects was related neither to divalent cation binding nor to the divalent cation-induced association of the protein.     

Calcium-induced formation of chlorophyll b and light-harvesting chlorophyll a/b-protein complex in cucumber cotyledons in the dark

Dark-grown cucumber seedlings were exposed to intermittent light (2 min light and 98 min dark) and then cotyledons were incubated with 50 mM CaCl₂ in the dark. Chlorophyll (Chl) a was selectively accumulated under intermittent light and Chl b was accumulated during the subsequent dark incubation with CaCl₂. The change in chlorophyll-protein complexes during Chl b accumulation induced by CaCl₂ in the dark was investigated by SDS-polyacrylamide gel electrophoresis. Chlorophyll-protein complex I and free chlorophyll were major chlorophyll-containing bands of the cotyledons intermittently illuminated 10 times. When these cotyledons were incubated with CaCl₂ in the dark, the light-harvesting Chl a/b-protein complex was formed. When the number of intermittent illumination periods was extended to 55, small amounts of Chl b and light-harvesting Chl a/b-protein complex were recognized at the end of intermittent light treatment, and these two pigments were further increased during the subsequent incubation of the cotyledons with CaCl₂ in the dark compared to water controls.     

Exciton interaction among chlorophyll molecules in bacteriochlorophyll a proteins and bacteriochlorophyll a reaction center complexes from green bacteria

Absorption and CD spectra of bacteriochlorophyll a proteins and bacteriochlorophyll a reaction center complexes from two strains of Chlorobium limicola were recorded at 77 °K. Visual inspection showed that the Q_y-band of chlorophyll in either protein was split into at least five components. Analysis of the spectra in terms of asymmetric Gaussian component pairs by means of computer program GAMET showed that six components are necessary to fit the spectra from strain 2K. These six components are ascribed to an exciton interaction between the seven bacteriochlorophyll a molecules in each subunit. The clear difference between the exciton splitting in the two bacteriochlorophyll a proteins shows that the arrangement of the chlorophyll molecules in each subunit must be slightly different.

The spectra for the bacteriochlorophyll a reaction center complexes have a component at 834 nm (absorption) and 832 nm (CD) which does not appear in the spectra of the bacteriochlorophyll a proteins. The new component is ascribed to a reaction center complex which is combined with bacteriochlorophyll a proteins to form the bacteriochlorophyll a reaction center complex. The complete absorption (or CD) spectrum for a given bacteriochlorophyll a reaction center complex can be described to a first approximation in terms of the absorption (or CD) spectrum for the corresponding bacteriochlorophyll a protein plus the new component ascribed to the reaction center complex.     

Thylakoid polypeptide composition and light-independent phosphorylation of the chlorophyll a,b-protein in Prochloron, a prokaryote exhibiting oxygenic photosynthesis

Thylakoids of the prokaryote Prochloron, present as a symbiont in ascidians isolated from the Red Sea at Eilat (Israel), showed polypeptide electrophoretic patterns comparable to those of thylakoids from eukaryotic oxygen-evolving organisms. Low temperature, fluorescence spectroscopy of Prochloron, having a chlorophyll a/b ratio of 3.8–5, and frozen in situ, demonstrated the presence of Photosystem II chlorophyll-protein complex emitting at 686 and 696 nm, as well as the emission band of Photosystem I at 720 nm which was so far not observed in Prochloron species. The latter emission was absent, if the cells or thylakoids were isolated prior to freezing. Energy transfer from chlorophyll b to chlorophyll a could be demonstrated to occur in vivo. The chlorophyll a,b-protein complex of Photosystem II, isolated by non-denaturing polyacrylamide gel electrophoresis, contained one major polypeptide of 34 kDa. The polypeptide was phosphorylated in vitro by a membrane-bound protein kinase which was not stimulated by light. A light-independent protein kinase activity was also found in isolated thylakoids of another prokaryote, the cyanophyte Fremyella diplosiphon. State I–State II transition could not be demonstrated in Prochloron by measurements of modulated fluorescence intensity in situ. We suggest that the presence of a light-independent thylakoid protein kinase of Prochloron, collected in the Red Sea at not less than 30 m depth, might be the result of an evolutionary process whereby this organism has adapted to an environment in which light, absorbed preferentially by Photosystem II, prevails.     

Excitation spectra for Photosystem I and Photosystem II in chloroplasts and the spectral characteristics of the distribution of quanta between the two photosystems

The parameters listed in the title were determined within the context of a model for the photochemical apparatus of photosynthesis.

The fluorescence of variable yield at 750 nm at −196 °C is due to energy transfer from Photosystem II to Photosystem I. Fluorescence excitation spectra were measured at −196 °C at the minimum, F_O, level and the maximum, F_M, level of the emission at 750 nm. The difference spectrum, F_M–F_O, which represents the excitation spectrum for F_V is presented as a pure Photosystem II excitation spectrum. This spectrum shows a maximum at 677 nm, attributable to the antenna chlorophyll a of Photosystem II units, with a shoulder at 670 nm and a smaller maximum at 650 nm, presumably due to chlorophyll a and chlorophyll b of the light-harvesting chlorophyll complex.

Fluorescence at the F_O level at 750 nm can be considered in two parts; one part due to the fraction of absorbed quanta, , which excites Photosystem I more-or-less directly and another part due to energy transfer from Photosystem II to Photosystem I. The latter contribution can be estimated from the ratio of F_O/F_V measured at 692 nm and the extent of F_V at 750 nm. According to this procedure the excitation spectrum of Photosystem I at −196 °C was determined by subtracting 1/3 of the excitation spectrum of F_V at 750 nm from the excitation spectrum of F_O at 750 nm. The spectrum shows a relatively sharp maximum at 681 nm due to the antenna chlorophyll a of Photosystem I units with probably some energy transfer from the light-harvesting chlorophyll complex.

The wavelength dependence of was determined from fluorescence measurements at 692 and 750 nm at −196 °C. is constant to within a few percent from 400 to 680 nm, the maximum deviation being at 515 nm where shows a broad maximum increasing from 0.30 to 0.34. At wavelengths between 680 and 700 nm, increases to unity as Photosystem I becomes the dominant absorber in the photochemical apparatus.     

Growth and mortality rates of phytoplankton in the northwestern North Pacific estimated by the dilution method and HPLC pigment analysis

Pigment-based growth rates of phytoplankton and mortality rates due to microzooplankton grazing were estimated using a dilution method combined with high-performance liquid chromatography (HPLC) pigment analysis in the northwestern North Pacific in autumn 1998. The dilution experiments were conducted at different hydrographic stations in both colder and warmer water masses. No significant difference was found between the growth rate of the phytoplankton community (0.38–0.70 day⁻¹; estimated by chlorophyll a) at the colder and warmer water stations, while the mortality rate (0.15–0.88 day⁻¹; estimated by chlorophyll a) tended to be higher at warmer water stations. The combination of estimates of daily chlorophyll a production and particulate organic carbon (POC) production enabled us to assess the carbon to chlorophyll a ratio (C/chl a) of “new” organic matter produced by living phytoplankton. The method provided an implicit value of the C/chl a of in situ living phytoplankton. The rate estimates from taxon-specific pigments suggested a possibility that chlorophyll b-containing green algae were grazed preferentially by microzooplankton during their active growth, and the standing stock of green algae was more strictly controlled by micrograzer than other algal groups such as diatoms. This result is one possible explanation for the fact that blooms of green algae have not been reported in the open ocean, in contrast with diatoms.     

Photochemical systems in mesophyll and bundle sheath chloroplasts of C4 plants

1. The agranal bundle sheath chloroplasts of Sorghum bicolor possess very low Photosystem II activity compared with the grana-containing mesophyll chloroplasts.

2. Sorghum mesophyll chloroplasts have a chlorophyll (chl) and carotenoid composition similar to that of spinach chloroplasts. In contrast, the sorghum bundle sheath chloroplasts have a higher chl a/chl b ratio; they are enriched in β-carotene and contain relatively less xanthophylls as compared to sorghum mesophyll or spinach chloroplasts.

3. Sorghum mesophyll chloroplasts with 1 cytochrome f, 2 cytochrome b₆ and 2 cytochrome b-559 per 430 chlorophylls have a cytochrome composition similar to spinach chloroplasts. Sorghum bundle sheath chloroplasts contain cytochrome f and cytochrome b₆ in the same molar ratios as for the mesophyll chloroplasts, but cytochrome b-559 is barely detectable.

4. The chl/P700 ratios of mesophyll chloroplasts of S. bicolor and mesophyll and bundle sheath chloroplasts of Atriplex spongiosa are similar to that of spinach chloroplasts suggesting that these chloroplasts possess an identical photosynthetic unit size to that of spinach. The agranal bundle sheath chloroplasts of S. bicolor possess a photosynthetic unit which contains only about half as many chlorophyll molecules per P700 as found in the grana-containing chloroplasts.

5. The similarity of the composition of the bundle sheath chloroplasts of S. bicolor with that of the Photosystem I subchloroplast fragments, together with their smaller photosynthetic unit and low Photosystem II activities suggests that these chloroplasts are highly deficient in the pigment assemblies of Photosystem II.     

Bacteriochlorophyll a-protein interactions in a complex from Prosthecochloris aestuarii. A Resonance Raman study

Raman spectra of bacteriochlorophyll a (BChl) bound to the soluble protein complex from Prosthecochloris aestuarii have been obtained at low temperature, using the resonance effect on their Q_x for Soret electronic bands. These spectra show that the acetyl carbonyls of at least four of the seven molecules bound to the monomer subunit of the complex and the ketone carbonyls of at least five of them are oriented close to the mean plane of the conjugated part of the dihydrophorbin macrocycle. Up to three bacteriochlorophyll molecules may have their ketone carbonyls free from hydrogen-bonding and up to two may have their acetyl carbonyls similarly free. Several of the binding sites of the remaining conjugated carbonyls are probably the same as those binding the conjugated carbonyls of bacteriochlorophyll (and of bacteriopheophytin) in reaction centers and in antenna structures of purple bacteria and as those binding chlorophyll in the antenna of higher plants and algae. The present resonance Raman spectra confirm that the magnesium atoms of most of the seven bacteriochlorophylls are pentacoordinated. They also show that polarisation effects from their local environments induce changes in the groundstate structures of the dihydrophorbin skeletons of the complexed molecules with respect to those of isolated, monomeric bacteriochlorophyll. These changes are quasi-identical for the seven molecules. These environmental effects predominate over any structural change brought about by intermolecular bonding of the conjugated carbonyls or of the magnesium atoms. The dihydrophorbin rings of the seven molecules thus appear to be immersed in a nearly homogeneous medium of low permittivity, although specific van der Waals interactions may polarise the free carbonyls to quite different extents. The possible implications of these observations on the interpretation of the electronic spectrum of the set of complexed bacteriochlorophylls are discussed.     

紫茉莉开花过程中花色和花色素的变化规律

紫茉莉是我国广泛分布的庭院花卉之一,具有丰富的花色。但不同花色紫茉莉在开花过程中的花色变化规律及其呈色机制还不清楚。以紫红色、黄色和白色紫茉莉为研究对象,分别通过色差仪测定法和紫外-可见分光光度法测定了不同开花时期不同花色紫茉莉花色表型及各类色素含量,探讨了其花色和色素变化规律,揭示其呈色机制。结果表明,从花蕾期到盛开期,紫红色紫茉莉花冠由淡绿色转变为紫红色,明度L^*值和色相b^*值减小,而色相a^*值、色度C^*值和色度角h值增大,叶绿素含量逐渐下降,类胡萝卜素、花色素苷和总黄酮含量逐渐升高;黄色紫茉莉花冠由淡绿色转变为黄色,盛开期具有最高的色度C^*值、色相a^*值和b^*值,整个开花过程具有较稳定的叶绿素和总黄酮含量,同时具有较高的类胡萝卜素含量;白色紫茉莉花冠由淡绿色转变为白色,过渡期具有最高的明度L^*值、色度C^*值、色相a^*值和b^*值,整个开花过程花色素苷和总黄酮含量较低,但随着开花进程逐渐升高,而类胡萝卜素含量稳定,过渡期总叶绿素含量显著低于其他2个时期。可见,不同花色紫茉莉开花过程中花色变化规律存在差异,而其差异性与其相应的色素成分变化密切相关。     

Monomolecular films of bacteriochlorophyll and derivatives at an air-water interface: Surface and spectral properties

Monomolecular films of bacteriochlorophyll, bacteriopheophytin and 2-desvinyl-2-acetyl chlorophyll a were prepared and studied on aqueous subphases containing pH 7.8 buffer and 4·10⁻⁴ M ascorbate. These monolayers are mechanically stable in the dark and light at 15 °C. at surface pressures below about 18 dynes/cm the slope of the surface isotherm of bacteriochlorophyll is steeper than at pressures greater than 18 dynes/cm. The surface dipole moments of bacteriochlorophyll are less than half that reported for chlorophyll a. Compression of bacteriochlorophyll or bacteriopheophytin monolayers result in changes of their absorption spectra.

Compression of bacteriochlorophyll monolayers to 18 dynes/cm results in a shift of the pigment's red peak from 787 to 749 nm as well as the appearance of a new absorption maximum at 896 nm. Continued compression to 24 dynes/cm results in a slight decrease in peak height of the 794-nm maximum and further increase in the absorbance of the 896-nm maximum. With bacteriopheophytin the red maximum at 760 nm starts to shift when the film is compressed to a surface pressure of only 2 dynes/cm; further compression yields a new absorption maximum at 846 nm. Compression of a film of 2-desvinyl-2-acetyl chlorophyll a results in only a 10-nm shift of the absorption maximum at 690 nm.

An orientation of bacteriochlorophyll at an air-water interface is proposed that is different from that for chlorophyll a. Like chlorophyll a bacteriochlorophyll monolayers are closely packed, but different in that bacteriochlorophyll allows greater interaction between pigment molecules. In compressed monolayers bacteriochlorophyll appears to aggregate differently than in other model systems.     

The dimerization of protochlorophyll pigments in non-polar solvents

The infrared, visible and nuclear magnetic resonance spectra of protochlorophyll a and vinylprotochlorophyll a in dry non-polar solvents (carbon tetrachloride, chloroform, cyclohexane) are presented and interpreted in terms of dimer interaction.

The infrared spectra in the 1600–1800 cm⁻¹ region clearly show the existence of a coordination interaction between the C-9 ketone oxygen function of one molecule and the central magnesium atom of another molecule. Infrared spectra in the OH stretching region (3200–3800 cm⁻¹) provide a valuable test of the water content in the samples.

The analysis of the absorption and circular dichroism spectra of protochlorophyll a and vinylprotochlorophyll a in carbon tetrachloride demonstrates the existence of a monomer-dimer equilibrium in the concentration range from 10⁻⁶ to 5 · 10⁻⁴ M. The dimerization constants are (6±2) · 10⁵ 1 · M⁻¹ for protochlorophyll a and (4.5±2) · 10⁵ 1 · M⁻¹ for vinylprotochlorophyll a at 20 °C. The deconvolution of visible spectra in the red region has been performed in order to obtain quantitative information on the dimer structure. Two models involving a parallel or a perpendicular arrangement of the associated molecules are considered.

From ¹H NMR spectra, it appears that the region of overlap occurs near ring V, in agreement with the interpretation of the infrared spectra.     

Structure and organization of System II photosynthetic units during the greening of a dark-grown Chlorella mutant

The formation and the organization of Photosystem II photosynthetic units during the greening of a dark-grown Chlorella vulgaris, mutant 5/520, have been investigated by analysing the kinetics of the “activation” of oxygen evolution and of the fluorescence induction.

1. 1. The existence during the early stages of the greening of a stationary photosynthesis demonstrates the presence of active Photosystem II at these initial stages, which are integrated in a functional whole, leading to overall photosynthesis.

2. 2. The rise-time of oxygen evolution has been measured using far-red and green light in order to estimate the relative number of chlorophylls per unit. The amount of chlorophyll a remains relatively constant during the greening, while the progressive addition of chlorophyll b causes the size of the units to increase approx. 2-fold.

3. 3. The induction kinetics of the fluorescence are exponential during the early phases of greening and later become distinctly sigmoidal; this suggests that the first units synthesized on the surface of the membrane are isolated from each other by obstacles preventing electronic excitation transfers and that such obstacles which might correspond to some distance between such units, can disappear at later stages, allowing energy transfers to occur.

These observations suggest that the Photosystem II units represent organized functional entities. They apparently consist of a relatively constant number of chlorophyll a molecules, which during the greening is complemented progressively by the addition of chlorophyll b.     

Respiratory cytochromes of Euglena gracilis

1. 1. Difference spectra of whole cells and of a particulate fraction of a streptomycin-bleached strain of Euglena gracilis showed the presence of a b-type cytochrome, cytochrome b (561 Euglena), and an a-type cytochrome, cytochrome a-type (609 Euglena). The cytochromes were characterized by pyridine hemochromogen formation and were found associated with a particulate fraction enriched with mitochondria.

2. 2. Both b-type and a-type cytochromes were reduced by succinate, oxidized by oxygen and reacted with a soluble c-type cytochrome, cytochrome c-type (556 Euglena), in reversible oxidation-reduction reactions. The steady-state level of reduction for each cytochrome was 92, 22 and 5% of the anaerobic level for the b-type, c-type and a-type cytochrome, respectively.

3. 3. Oxidation of c-type and a-type cytochromes was completely inhibited by cyanide, although respiration of a particulate fraction was only 60% inhibited by the same concentration of cyanide. Antimycin A inhibited respiration by up to 70% but completely inhibited reduction of the c-type cytochrome.

4. 4. The data suggest that electron transfer in the respiratory pathway of Euglena involves the b-, c- and a-type cytochrome in a direct sequence. The cyanide and antimycin A-insensitive oxidation pathway is considered to involve a more direct oxidation of the b-type cytochrome.

Structure of the primary electron donor in photosystem I: a resonance Raman study

Low-temperature resonance Raman (RR) spectra have been obtained at resonance with the Soret transition of chlorophyll a in photosystem I particles containing large amounts either of the triplet state of P700 or of its radical cation state. Subtracting these spectra from those of resting reaction centers yielded RR spectra of P700 in its neutral, ground state. These spectra arise from two distinct chlorophyll a molecules differing by the strengths of the bonding interactions assumed by their keto carbonyl groups, the stretching frequencies of which are found at 1655 and 1675 cm-1. The present results rule out previous hypotheses that P700 might have consisted of a single, chemically modified chlorophyll a molecule. Neither of the bonding interactions assumed by the keto carbonyls of the P700 chlorophylls most probably involves chlorophyll-chlorophyll bridging through water molecules, as surmised in the so-called special pair models, but likely consists of H bonds with distinct protein sites. The magnesium atoms of the two P700 chlorophylls are 5-coordinated. Hence, the structural model of P700 provided by the present data is qualitatively the same, in terms of bonding interactions, as that currently accepted for the bacterial primary donor.