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Biomedical subjects

A Melis

Publications and source records attributed to A Melis.

At least 73 records · Page 4Linked to original sources
PubMed1985

Characterization of a cyanobacterial photosystem I complex.

A simple procedure is described for the preparation of photosystem I (PSI) particles from Triton X-100-solubilized thylakoid membranes of the unicellular cyanobacterium Synechococcus 6301. The purified PSI complex contained the full complement of antenna chlorophylls, 130 +/- 5/P700, displayed the electron paramagnetic resonance signals characteristic of iron-sulfur centers X, A, and B, and had a protein/chlorophyll ratio of 2.9. Determination of the polypeptide composition, utilizing a uniformly 14C-labeled complex, showed that it contained polypeptides of 70, 18, 17.7, 16, and 10 kDa, in a molar ratio of 4.0:0.7:1.0:0.5:1.6. The relative amount of the lower molecular weight polypeptides showed progressive decrease with increase in Triton X-100 concentration and time of exposure to detergent. Consequently, it is proposed that in vivo the composition of the complex is [70 kDa]4 [18 kDa]1 [17.7 kDa]1 [16 kDa]1 [10 kDa]2. Relative to 130 mol of chlorophyll a, the PSI complex contained 16 mol of carotenoids, 13.7 +/- 1.0 g atoms of Fe, and 12.2 +/- 1.1 g atoms of labile sulfide. The properties of complexes fully depleted of the low-molecular weight polypeptides by treatment with sodium dodecyl sulfate or with proteinase K are also described.

Chlorophyll↗
PubMed1985

Photochemical Apparatus Organization in the Chloroplasts of Two Beta vulgaris Genotypes.

The composition and structural organization of thylakoid membranes of a low chlorophyll mutant of Beta vulgaris was investigated using spectroscopic, kinetic and electrophoretic techniques. The data obtained were compared with those of a standard F1 hybrid of the same species. The mutant was depleted in chlorophyll b relative to the hybrid and it had a higher photosystem II/photosystem I reaction center (Q/P(700)) ratio and a smaller functional chlorophyll antenna size. Analysis of thylakoid membranes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the mutant lacked a portion of the chlorophyll a/b light-harvesting complex but was enriched in the photosystem II reaction center chlorophyll protein complex. Comparison of functional antenna sizes and of photosystem stoichiometries determined electrophoretically were in good agreement with those determined spectroscopically. Both approaches indicated that about 30% of the total chlorophyll was associated with photosystem I and about 70% with photosystem II. A greater proportion of photosystem II(beta) was detected in the mutant. The results suggest that a higher photosystem II to photosystem I ratio in the sugar beet mutant has apparently compensated for the smaller photosystem II chlorophyll light-harvesting antenna in its chloroplasts. Moreover, a lack of chlorophyll a/b light-harvesting complex correlates with the abundance of photosystem II(beta). It is proposed that a developmental relationship exists between the two types of photosystem II where photosystem II(beta) is a precursor form of photosystem II(alpha) occurring prior to the addition of the chlorophyll a/b light-harvesting complex and grana formation.

Journal Article↗
PubMed1984

Light regulation of photosynthetic membrane structure, organization, and function.

The light environment during plant growth determines the structural and functional properties of higher plant chloroplasts, thus revealing a dynamically regulated developmental system. Pisum sativum plants growing under intermittent illumination showed chloroplasts with fully functional photosystem (PS) II and PSI reaction centers that lacked the peripheral chlorophyll (Ch1) a/b and Ch1 a light-harvesting complexes (LHC), respectively. The results suggest a light flux differential threshold regulation in the biosynthesis of the photosystem core and peripheral antenna complexes. Sun-adapted species and plants growing under far-red-depleted illumination showed grana stacks composed of few (3-5) thylakoids connected with long intergrana (stroma) thylakoids. They had a PSII /PSI reaction center ratio in the range 1.3-1.9. Shade-adapted species and plants growing under far-red-enriched illumination showed large grana stacks composed of several thylakoids, often extending across the entire chloroplast body, and short intergrana stroma thylakoids. They had a higher PSII /PSI reaction center ratio, in the range of 2.2-4.0. Thus, the relative extent of grana and stroma thylakoid formation corresponds with the relative amounts of PSII and PSI in the chloroplast, respectively. The structural and functional adaptation of the photosynthetic membrane system in response to the quality of illumination involves mainly a control on the rate of PSII and PSI complex biosynthesis.

Chlorophyll↗
PubMed1984

Photochemical Apparatus Organization in Anacystis nidulans (Cyanophyceae) : Effect of CO(2) Concentration during Cell Growth.

Anacystis nidulans cells grown under high (3%) CO(2) partial pressure have greater phycocyanin to chlorophyll ratio (Phc/Chl) relative to cells grown under low (0.2%) CO(2) tension (Eley (1971) Plant Cell Physiol 12: 311-316). Absorbance difference spectrophotometry of A. nidulans thylakoid membranes in the ultraviolet (DeltaA(320)) and red (DeltaA(700)) regions of the spectrum reveal photosystem II/photosystem I (PSII/PSI) reaction center ratio (RCII/RCI) changes that parallel those of Phc/Chl. For cells growing under 3% CO(2), the Phc/Chl ratio was 0.48 and RCII/RCI = 0.40. At 0.2% CO(2), Phc/Chl = 0.38 and RCII/RCI = 0.24. Excitation of intact cells at 620 nm sensitized RCII at a rate approximately 20 times faster than that of RCI, suggesting that Phc excitation is delivered to RCII only. In the presence of DCMU, excitation at 620 nm induced single exponential RCII photoconversion kinetics, suggesting a one-to-one structural-functional correspondance between phycobilisome and PSII complex in the thylakoid membrane. Therefore, phycobilisomes may serve as microscopic markers for the presence of PSII in the photosynthetic membrane of A. nidulans. Neither the size of individual phycobilisomes nor the Chl light-harvesting antenna of PSI changed under the two different CO(2) tensions during cell growth. Our results are compatible with the hypothesis that, at low CO(2) concentrations, the greater relative amounts of PSI present may facilitate greater rates of ATP synthesis via cyclic electron flow. The additional ATP may be required for the active uptake of CO(2) under such conditions.

Journal Article↗
PubMed1984

Photosystem electron-transport capacity and light-harvesting antenna size in maize chloroplasts.

Spectrophotometric and kinetic measurements were applied to yield photosystem (PS) stoichiometries and the functional antenna size of PSI, PSII(alpha), and PSII(beta) in Zea mays chloroplasts in situ. Concentrations of PSII and PSI reaction centers were determined from the amplitude of the light-induced absorbance change at 320 and 700 nm, which reflect the photoreduction of the primary electron acceptor Q of PSII and the photooxidation of the reaction center P700 of PSI, respectively. Determination of the functional chlorophyll antenna size (N) for each photosystem was obtained from the measurement of the rate of light absorption by the respective reaction center. Under the experimental conditions employed, the rate of light absorption by each reaction center was directly proportional to the number of light-harvesting chlorophyll molecules associated with the respective photosystem. We determined N(P700) = 195, N(alpha) = 230, N(beta) = 50 for the number of chlorophyll molecules in the light-harvesting antenna of PSI, PSII(alpha), and PSII(beta), respectively. The above values were used to estimate the PSII/PSI electron-transport capacity ratio (C) in maize chloroplasts. In mesophyll chloroplasts C > 1.4, indicating that, under green actinic excitation when Chl a and Chl b molecules absorb nearly equal amounts of excitation, PSII has a capacity to turn over electrons faster than PSI. In bundle sheath chloroplasts C < 1, suggesting that such chloroplasts are not optimally poised for linear electron transport and reductant generation.

Journal Article↗
PubMed1983

Localization of photosynthetic electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays.

The organization of the electron transport components in mesophyll and bundle sheath chloroplasts of Zea mays was investigated. Grana-containing mesophyll chloroplasts (chlorophyll a to chlorophyll b ratio of about 3.0) possessed the full complement of the various electron transport components, comparable to chloroplasts from C3 plants. Agranal bundle sheath chloroplasts (Chl a/Chl b greater than 5.0) contained the full complement of photosystem (PS) I and of cytochrome (cyt) f but lacked a major portion of PS II and its associated Chl a/b light-harvesting complex (LHC), and most of the cyt b559. The kinetic analysis of system I photoactivity revealed that the functional photosynthetic unit size of PS I was unchanged and identical in mesophyll and bundle sheath chloroplasts. The results suggest that PS I is contained in stroma-exposed thylakoids and that it does not receive excitation energy from the Chl a/b LHC present in the grana. A stoichiometric parity between PS I and cyt f in mesophyll and bundle sheath chloroplasts indicates that biosynthetic and functional properties of cyt f and P700 are closely coordinated. Thus, it is likely that both cyt f and P700 are located in the membrane of the intergrana thylakoids only. The kinetic analysis of PS II photoactivity revealed the absence of PS II alpha from the bundle sheath chloroplasts and helped identify the small complement of system II in bundle sheath chloroplasts as PS II beta. The distribution of the main electron transport components in grana and stroma thylakoids is presented in a model of the higher plant chloroplast membrane system.

Binding Sites↗
PubMed1983

Localization of different photosystems in separate regions of chloroplast membranes.

The stoichiometric amounts and the photoactivity kinetics of photosystem I (PSI) and of the alpha and beta components of photosystem II (PSII(alpha) and PSII(beta)) were compared in spinach chloroplast membrane (thylakoid) fractions derived from appressed and nonappressed regions. Stroma-exposed thylakoid fractions from the nonappressed regions were isolated by differential centrifugation following a mechanical press treatment of the chloroplasts. Thylakoid vesicles derived mainly from the appressed membranes of grana were isolated by the aqueous polymer two-phase partition method. Stroma-exposed thylakoids were found to have a chlorophyll a/chlorophyll b ratio of 6.0 and a PSII(beta)/PSI reaction center ratio of 0.3. Kinetic analysis of system II photoactivity revealed the absence of PSII(alpha) from stroma-exposed thylakoids. The photoactivity of system I in stroma-exposed thylakoids showed a single kinetic component identical to that of unfractionated chloroplasts, suggesting that PSI does not receive excitation energy from the PSII-chlorophyll ab light-harvesting complex. Thus, stroma-exposed thylakoids are significantly enriched in both PSI and PSII(beta). Inside-out vesicles from the appressed membranes of grana-partition regions had a chlorophyll a/chlorophyll b ratio of 2.0 and a PSII/PSI reaction center ratio of 10.0. The photoactivity of system II showed the membranes of the grana-partition regions to be significantly enriched in PSII(alpha). We conclude that PSII(alpha) is exclusively located in the membranes of the grana partitions while PSII(beta) and PSI are located in stroma-exposed thylakoids. The low PSI reaction center (P700) content of vesicles derived from grana partitions and the kinetic homogeneity of the PSI complex suggest total exclusion of P700 as a functional component in the membrane of the grana-partition region.

Journal Article↗
PubMed1982

Chlorophyll a Fluorescence Transients in Mesophyll and Guard Cells : MODULATION OF GUARD CELL PHOTOPHOSPHORYLATION BY CO(2).

Chlorophyll fluorescence transients from mesophyll and guard cell chloroplasts of variegated leaves from Chlorophytum comosum were compared using high resolution fluorescence spectroscopy. Like their mesophyll counterparts, guard cell chloroplasts showed the OPS fluorescence transient indicating the operation of the linear electron transport and the possible generation of NADPH in these organelles. They also showed a slow fluorescence yield decrease, equivalent to the MT transition in mesophyll, suggesting the formation of the high energy state and photophosphorylation. Unlike the mesophyll chloroplasts, the fluorescence from guard cell chloroplasts lacked the increment of the SM transition, indicating that the two types of chloroplasts have some metabolic differences. The presence of CO(2) (supplied as bicarbonate, pH 6.7) specifically inhibited the MT-equivalent transition while its absence accelerated it. These observations constitute the first specific evidence of a guard cell chloroplast response to CO(2). Control of photosynthetic ATP levels in the guard cell cytoplasm by CO(2) may provide a mechanism regulating the availability of high energy equivalents at the guard cell plasmalemma, thus affecting stomatal opening.

Journal Article↗
PubMed1981

Fluorescence Properties of Guard Cell Chloroplasts: EVIDENCE FOR LINEAR ELECTRON TRANSPORT AND LIGHT-HARVESTING PIGMENTS OF PHOTOSYSTEMS I AND II.

The presence of chloroplasts in guard cells from leaf epidermis, coleoptile, flowers, and albino portions of variegated leaves was established by incident fluorescence microscopy, thus confirming the notion that guard cell chloroplasts are remarkably conserved. Room temperature emission spectra from a few chloroplasts in a single guard cell of Vicia faba showed one major peak at around 683 nanometers. Low-temperature (77 K) emission spectra from peels of albino portions of Chlorophytum comosum leaves and from mesophyll chloroplasts of green parts of the same leaves showed major peaks at around 687 and 733 nanometers, peaks usually attributed to photosystem II and photosystem I pigment systems, respectively. Spectra of peels of V. faba leaves showed similar peaks. However, fluorescence microscopy revealed that the Vicia peels, as well as those from Allium cepa and Tulipa sp., were contaminated with non-guard cell chloroplasts which were practically undetectable under bright field illumination. These observations pose restrictions on the use of epidermal peels as a source of isolated guard cell chloroplasts. Studies on the 3-(3,4-dichlorophenyl)-1,1-dimethylurea-sensitive variable fluorescence kinetics of uncontaminated epidermal peels of C. comosum indicated that guard cell chloroplasts operate a normal, photosystem II-dependent, linear electron transport. The above properties in combination with their reported inability to fix CO(2) photosynthetically may render the guard cell chloroplasts optimally suited to supply the reducing and high-energy phosphate equivalents needed to sustain active ion transport during stomatal opening in daylight.

Journal Article↗
PubMed1980

The relative absorption cross-sections of photosystem I and photosystem II in chloroplasts from three types of Nicotiana tabacum.

In the present study we used three types of Nicotiana tabacum, cv John William's Broad Leaf (the wild type and two mutants, the yellow-green Su/su and the yellow Su/su var. Aurea) in order to correlat functional properties of Photosystem II and Photosystem I with the structural organization of their chloroplasts. The effective absorption cross-section of Photosystem II and Photosystem I centers was measured by means of the rate constant of their photoconversion under light-limiting conditions. In agreement with earlier results (Okabe, K., Schmid, G.H. and Straub, J. (1977) Plant Physiol. 60, 150--156) the photosynthetic unit size for both System II and System I in the two mutants was considerably smaller as compared to the wild type. We observed biphasic kinetics in the photoconversion of System II in all three types of N. tabacum. However, the photoconversion of System I occurred with monophasic and exponential kinetics. Under our experimental conditions, the effective cross-section of Photosystem I was comparable to that of the fast System II component (alpha centers). The relative amplitude of the slow System II component (beta centers) varied between 30% in the wild type to 70% in the Su/su var. Aurea mutant. The increased fraction of beta centers is correlated with the decreased fraction of appressed photosynthetic membranes in the chloroplasts of the two mutants. As a working hypothesis, it is suggested that beta centers are located on photosynthetic membranes directly exposed to the stroma medium.

Chloroplasts↗
PubMed1980

Stoichiometry of system I and system II reaction centers and of plastoquinone in different photosynthetic membranes.

The concentrations of photochemical centers and of plastoquinone were measured in several kinds of photosynthetic membranes by optical difference spectroscopy. Photosystem I reaction centers were measured from the light-induced absorbance change at 700 nm (oxidation of the primary electron donor, P700). Photosystem II reaction centers were estimated from the light-induced absorbance change at 325 nm (reduction of the primary electron acceptor, Q). Spinach chloroplasts and membrane fractions obtained by French press treatment, mature and developing pea chloroplasts, and blue-green algal membranes were investigated. No loss of primary photochemical activity occurred during fractionation of the chloroplasts. The results indicated a large variability in the ratio of system II to system I reaction centers (from 0.43 to 3.3) in different photosynthetic membranes. Oxygen-evolving plants may change the ratio of their photosystems in response to environmental light conditions. The amount of photoreducible plastoquinone was also measured at 263 nm. In spinach chloroplasts, seven to eight plastoquinone molecules were found per reaction center of system II. Most of the plastoquinone pool was associated with the grana. However, the ratio of chemically determined plastoquinone to chlorophyll was similar in the grana and stroma thylakoids.

Journal Article↗
PubMed1979

The kinetic relationship between the C-550 absorbance change, the reduction of Q(delta A320) and the variable fluorescence yield change in chloroplasts at room temperature.

The light minus dark difference spectrum and the kinetics of the indicator pigment C-550 have been measured at room temperature in isolate, envelope-free chloroplasts in the presence of 3-(3' ,4'-dichlorophenyl)-1,1-dimethylurea (DCMU). The C-550 spectrum indicates a band shift with peaks at 540 and 550 nm and has an isobestic point at 545 nm. On the assumption of 400 chlorophyll molecules per electron transfer chain the differentaial extinction coefficient delta epsilon (540-550) is calculated to be approximately 5 mM-1 . CM-1. The kinetics of the C-550 absorbance change, occurring upin the onset of continuous illumination, are shown to be biphasic and strictly correlated with the kinetics of the complementary area measured from the fluorescence induction curve under identical cinditions and with those of the absorbance increase at 320 nm due to photoreduction of Q. The lighted-induced change in these three parameters can be described as a function of the variable fluorescence yield change occurring under the same conditions. Such functions are non-linear and reveal a heterogeneous dependence of the variable fluorescence yield on the fraction of closed System II reaction centers. It is concluded that for every molecule of the primary electron acceptor Q of Photosystem II that is photochemically reduced there corresponds an equivalent change in the absorbance of the indicator pigment C-550 and in the size of the complementary area. Ths, C-550 and area are two valid parameters for monitoring the primary photochemical activity of System II at the room temperature.

Chloroplasts↗
PubMed1977

Development of the Two Heterogeneous Photosystem II Units in Etiolated Bean Leaves.

The development of the photosystem II units in relation to the heterogeneity of their photochemical centers was studied in etiolated bean leaves (Phaseolus vulgaris var. red kidney) greened under continuous or intermittent light. The study was done in order to see whether grana are the loci of the units with the efficient photosystem II activity (alpha units), while the stroma thylakoids are the loci of the units with the less efficient photosystem II activity (beta units), as it has been proposed. In addition, the interrelations between alpha and beta centers have been investigated. It was found that the alpha and the beta centers of photosystem II were present in the first photosynthetic membranes irrespective of the mode of greening of the leaves. The magnitude of their respective photochemical rate constants, K'(alpha) and K(beta), increased with time in continuous light and it reached the steady-state values of the mature chloroplasts within 16 hours, while in intermittent light it remained smaller. The differentiation of the system II units in alpha and beta centers containing units is more evident under conditions of intermittent illumination, i.e. when the rate of chlorophyll biosynthesis is the limiting step for chloroplast development.It is concluded that the heterogeneity of the photochemical centers in system II is an endogenous property of the chloroplast lamellae. The alpha centers and the beta centers develop independently of each other from the beginning of the light-induced greening. They do not share the same pigment beds. The presence of grana, chlorophyll b, and the chlorophyll-protein complex II is not a prerequisite for the formation or development of the alpha centers. The formation of these centers precedes grana formation in greening plastids.

Journal Article↗