Vladimir A. Boichenko

Functional characteristics of chlorophyll d-predominating photosynthetic apparatus in intact cells of Acaryochloris marina

Functional organization of the photosynthetic apparatus in the unique chlorophyll d-predominating prokaryote, Acaryochloris marina, was studied using polarographic measurements of single-turnover flash yields, action spectra and optical cross sections for PS-specific reactions. O2 evolution was indicative of PS II activity, while reversible photoinhibition of respiratory O2 uptake under aerobic conditions in the presence of DCMU and H2 photoevolution by anaerobically adapted cells were the indicatives of PS I activity. O2 evolution in the cells upon single-turnover flashes followed the normal S-state cycle with a period-4 oscillation. Analysis of action spectra for the partial reactions of photosynthesis revealed that: (1) distinct spectral forms of Chl d are nonuniformly distributed between PS I and PS II, e.g. Chl d-695 and Chl d-735 are preferentially located in PS II and PS I, respectively; (2) a minor fraction of Chl a in the cells belongs mostly to PS II; (3) biliproteins transfer excitation energy both to PS II and, with a lower efficiency, PS I; (4) the efficiency of energy transfer from biliproteins to PS II depends on the light quality growth conditions and is larger in white light (WL)-grown cells compared to the red light (RL)-grown cells. Content of functional O2 evolving PS II centers decreases 2 times in the RL-grown cells relative to the WL-grown cells, whereas content of competent PS I centers involved in photoinhibition of respiration remains almost the same in both the cultures. The effective antenna size of PS I was estimated to be 80–90 Chl d including 3–10 molecules absorbing at 735 nm. The effective optical cross-section of PS II corresponded to 90–100 Chl d and, presumably, 4 Chl a + 2 Pheo a [Mimuro et al. (1999) Biochim Biophys Acta 1412: 37–46]. Optical cross-section measurements indicated that the functional PS II units of A. marina attach one rod of four hexameric units of biliproteins.

Action spectra and functional antenna sizes of Photosystems I and II in relation to the thylakoid membrane organization and pigment composition

The functional organization of competent photosynthetic units in developing thylakoids from intermittent-light grown pea as well as in the unstacked, stacked and phosphorylated stacked thylakoids from its mature chloroplasts was characterized by polarographic measurements of action spectra, reaction centre contents and optical cross-sections for PS I-mediated O2 uptake and PS II-mediated O2 evolution. The minimum antenna sizes of 60 and 37 chlorophyll a molecules for PS I and PS II, respectively, were determined in developing thylakoids with a ratio of Chl a/Chl b>50. In mature chloroplasts, the embedded light-harvesting chlorophyll a/b-binding (LHC) protein complexes increased the PS I and PS II effective antenna sizes by 3–6 times depending on the thylakoid membrane organization. In unstacked thylakoids, a randomization of PS I, PS II and LHC II led to the most uniform spectral distribution of light harvesting between the two photosystems but caused the maximal difference of their antenna sizes to be 370 and 100 Chls for the competent PS I and PS II units, respectively. Following the Mg2+-induced stacking of thylakoids, opposite complementary changes of the action spectra, antenna sizes and Chl a/Chl b ratios indicated a redistribution of a LHC II pool of ∼100 Chl ( a + b) molecules from PS I to PS II. Unlike to the stroma-exposed PS IIβ in unstacked thylakoids, the granal PS IIα units of ∼200 Chls demonstrated an additional 2-fold increase of the effective antenna size due to energy transfer within PS II dimers under strong background illumination, which closed >90% of reaction centres. Protein phosphorylation of the stacked thylakoids induced a significant inactivation of the O2-evolving PS II centres but did not cause complementary changes of the action spectra and antenna sizes of the competent PS I and PS II. In this case, light harvesting parameters of the O2-evolving PS II units were nearly unaffected, whereas the obvious relative increase of the PS I activity at 650 nm and its decrease at >700 nm both in the action spectrum and optical cross-section measurements might suggest a substitution of PS Iβ units in the O2-reducing fraction by another distinct fraction of α-type which in turn is not the same to PS Iα units in unstacked thylakoids.

Verification of the Z-scheme in Chlamydomonas mutants with Photosystem I deficiency

In conflict with the Z-scheme of photosynthesis, it has recently been reported [Greenbaum et al. Nature (1995) 376: 438–441; Lee et al. Science (1996) 273: 364–367] that Photosystem II can drive ferredoxin reduction and photoautotrophic growth in some mutants of Chlamydomonas lacking detectable Photosystem I reaction centre, P700. Using the same mutants, B4 and F8, here we report that action spectra and parameters of flash yields of different photoreactions show the operation in ferredoxin-dependent H2 photoproduction and CO2 fixation of a fraction (at least 5% compared to wild- type) of the only Photosystem I complexes.

Can Photosystem II be a photogenerator of low potential reductant for CO2 fixation and H2 photoevolution

The experimental grounds of the hypothesis of a single-light-reaction in oxygenic photosynthesis as suggested by Greenbaum et al. (Nature (1995) 376: 438–441) are critically discussed and a possible explanation of their data by Photosystem I contamination in the mutant utilized is argued.

Photosynthetic units of phototrophic organisms

Photoautotrophic organisms play a key role in the biosphere of the Earth, converting solar energy of the 350-1000 nm range into biochemically available form. In contemporary aquatic and terrestrial ecosystems, the dominating groups are the oxygen evolving cyanobacteria, algae, and higher plants. Anoxygenic phototrophic microorganisms occupy mainly ecological niches with extreme environmental conditions. Despite diverse evolution of all these taxonomic groups, their photosynthetic apparatus has a similar molecular design and identical principles of operation. This review covers recent data about features of the structural and functional organization of pigment–protein complexes of the basic types of photosynthetic units in prokaryotes and eukaryotes. A correspondence between the optical properties of various photosynthetic units and the natural light conditions is discussed.

Properties of mutant reaction centers of Rhodobacter sphaeroides with substitutions of histidine L153, the axial Mg2+ ligand of bacteriochlorophyll BA

Mutant reaction centers (RC) from Rhodobacter sphaeroides have been studied in which histidine L153, the axial ligand of the central Mg atom of bacteriochlorophyll BA molecule, was substituted by cysteine, methionine, tyrosine, or leucine. None of the mutations resulted in conversion of the bacteriochlorophyll BA to a bacteriopheophytin molecule. Isolated H(L153)C and H(L153)M RCs demonstrated spectral properties similar to those of the wild-type RC, indicating the ability of cysteine and methionine to serve as stable axial ligands of the Mg atom of bacteriochlorophyll BA. Because of instability of mutant H(L153)L and H(L153)Y RCs, their properties were studied without isolation of these complexes from the photosynthetic membranes. The most prominent effect of the mutations was observed with substitution of histidine by tyrosine. According to the spectral data and the results of pigment analysis, the BA molecule is missing in the H(L153)Y RC. Nevertheless, being associated with the photosynthetic membrane, this RC can accomplish photochemical charge separation with quantum yield of approximately 7% of that characteristic of the wild-type RC. Possible pathways of the primary electron transport in the H(L153)Y RC in absence of photochemically active chromophore are discussed.

Mass spectrometric study of Photosystem II heterogeneity in oxygen and nitrogen production: Effects of magnesium and of phosphorylation of pea thylakoids

Photosystem II (PS II) is capable of the oxidation of both water and hydroxylamine with the production of O2- and N2-production, respectively. The resulting changes in the partial pressure of the respective gases can be measured by an appropriate mass spectrometric set-up. Analysis of single turn-over flash saturation curves of O2- and N2-production has been performed to determine the relative optical cross sections of the competent PS II units and absolute amounts of their fractions in pea thylakoids. We studied the changes of these parameters upon Mg2+-induced transition of thylakoid membrane from unstacked to stacked configuration and upon protein phosphorylation of the stacked samples. The results showed a 2.5-fold increase of effective antenna size of PS II units competent in either O2- or N2-production after addition of 10 mM MgCl2 to cation-depleted thylakoids, which indicates a potential capability of both α- and β-units to carry out these alternative reactions. However, we observed a significant difference in the amounts of PS II units competent in O2- or N2-production, with a ratio of 1:4 in unstacked thylakoids, and reciprocal alterations in stacked ones. This represents an increase by about 20% and a 2-fold decrease of O2- and N2-evolving units, respectively, yielding a ratio of 1:1.5, which implies a heterogeneity of PS II with respect to these reactions, the capabilities of α- and β-units being distinct. The phosphorylation of stacked thylakoids did not essentially influence the antenna size of O2- and N2-evolving PS II units but caused opposite and reciprocal changes in their amounts, approximately 30% decrease and increase, respectively, to a ratio of 1:3. The relationship of the structure-function heterogeneity in PS II with implications for current models of photosynthetic regulation mechanisms is discussed.