Freeze-fracture analysis of thylakoid membranes and photosystem I and II enriched fractions from Phormidium laminosum

1986 ◽  
Vol 80 (1) ◽  
pp. 57-73
Author(s):  
R.E. Glick ◽  
R.E. Triemer ◽  
B.A. Zilinskas

Thylakoid membranes of the thermophilic cyanobacterium Phormidium laminosum have been fractionated into photosystem II and photosystem I particles. These fractions have been characterized by their partial electron transport activities, and biochemical and spectral properties. Exoplasmic fracture face and protoplasmic fracture face particles in the unfractionated thylakoid membranes were shown to correspond in size to particles in freeze-fractured photosystem II and photosystem I fractions, respectively. Differences between the histograms of the thylakoid membrane protoplasmic fracture face particles and the isolated photosystem I particles suggest that in addition to photosystem I complexes some of the particles on the thylakoid protoplasmic fracture face may be related to cytochrome b/f complexes, the hydrophobic component of the coupling factor, or respiratory complexes.

1977 ◽  
Vol 73 (2) ◽  
pp. 400-418 ◽  
Author(s):  
P A Armond ◽  
L A Staehelin ◽  
C J Arntzen

We have previously demonstrated (Armond, P. A., C. J. Arntzen, J.-M. Briantais, and C. Vernotte. 1976. Arch. Biochem. Biophys. 175:54-63; and Davis, D. J., P. A. Armond, E. L. Gross, and C. J. Arntzen. 1976. Arch. Biochem. Biophys. 175:64-70) that pea seedlings which were exposed to intermittent illumination contained incompletely developed chloroplasts. These plastids were photosynthetically competent, but did not contain grana. We now demonstrate that the incompletely developed plastids have a smaller photosynthetic unit size; this is primarily due to the absence of a major light-harvesting pigment-protein complex which is present in the mature membranes. Upon exposure of intermittent-light seedlings to continuous white light for periods up to 48 h, a ligh-harvesting chlorophyll-protein complex was inserted into the chloroplast membrane with a concomitant appearance of grana stacks and an increase in photosynthetic unit size. Plastid membranes from plants grown under intermediate light were examined by freeze-fracture electron microscopy. The membrane particles on both the outer (PF) and inner (EF) leaflets of the thylakoid membrane were found to be randomly distributed. The particle density of the PF fracture face was approx. four times that of the EF fracture face. While only small changes in particle density were observed during the greening process under continuous light, major changes in particle size were noted, particularly in the EF particles of stacked regions (EFs) of the chloroplast membrane. Both the changes in particle size and an observed aggregation of the EF particles into the newly stacked regions of the membrane were correlated with the insertion of light-harvesting pigment-protein into the membrane. Evidence is presented for identification of the EF particles as the morphological equivalent of a "complete" photosystem II complex, consisting of a phosochemically active "core" complex surrounded by discrete aggregates of the light-harvesting pigment protein. A model demonstrating the spatial relationships of photosystem I, photosystem II, and the light-harvesting complex in the chloroplast membrane is presented.


1988 ◽  
Vol 43 (5-6) ◽  
pp. 423-430 ◽  
Author(s):  
J. Bednarz ◽  
A. Radunz ◽  
G. H. Schmid

The lipids of photosystem II particles, of chloroplasts and leaves are compared in the variegated tobacco mutant NC 95. The mutant differs from other N. tabacum mutants by the phenomenon that it has variegated leaves with green and with yellow-green leaf patches. Chloroplasts from the green leaf areas exhibit photosystem II and photosystem I reactions and have a normal lamellar system with grana and intergrana regions. Chloroplasts from the yellow-green leaf areas, however, yield only photosystem I reactions and have only single stranded isolated thylakoids. Hence, this mutant offers the unique possibility to compare without the use of detergents within the same plant the lipid composition of photosystem II particles with that in intact chloroplasts, exhibiting either photosystem II and I reactions or those exhibiting exclusively photosystem I reactions. The lipids of photosystem II particles are composed of 37 % glycolipids, 4 % phospholipids, 5 % carotenoids and 54 % chlorophyll. Lipids of chloroplasts with grana stacking are composed of 75% glycolipids, 7 % phospholipids, 2 % carotenoids and 16% chlorophyll. Chloroplasts with single isolated thylakoids have a lipid composition consisting of 8 3 % glycolipids, 14% phospholipids and only 0.5% carotenoids and 2 % chlorophyll. The chloroplast lipid mixture is characterized in comparison to the respective leaf lipid mixture by a 16-17% higher glycolipid portion and by a 13-70% lower phospholipid content. The main difference in the lipid composition of photosystem I and II consists in the observation that chloroplasts active in only photosystem I contain more than double the amount of glycolipids and the 4-fold amount of phospholipids in comparison to photosystem II active preparations. The amount of monogalactolipid is even 3 times higher in chloroplasts active only in photosystem I when compared to those in photosystem II particles. In photosystem II particles phosphatidylethanolamine is completely lacking and phosphatidylglycerol and phosphatidylinositol occur only in traces. The fatty acids of the sulfolipid are by 45 % more saturated in the photosystem II particles and the digalactolipids of the photosystem II particles are by 28 % more saturated than in chloroplasts exhibiting photosystem I and II activity.


1986 ◽  
Vol 80 (1) ◽  
pp. 13-28
Author(s):  
B. Kachar ◽  
N.A. Christakis ◽  
T.S. Reese ◽  
N.J. Lane

Smooth septate junctions from the midgut of the cricket, Acheta, and the horseshoe crab, Limulus, as well as Hydra-type septate junctions from the epidermis of Hydra have been studied by freeze-fracture after direct freezing using the liquid helium-cooled copper block/slam freezing method. The exoplasmic fracture face at both types of septate junction exhibits rows of closely packed but irregularly shaped intramembrane particles. Complementary to these particle rows, on the protoplasmic fracture face, are sharply defined grooves with a periodic variation in depth and width that was conspicuous in Hydra but less well defined in arthropods. The closely packed, irregular particles on the exoplasmic faces could represent plastically deformed portions of transmembrane proteins pulled through the bilayer during freeze-fracture. On the basis of this interpretation, the grooves on the protoplasmic faces represent a confluence of the bilayer disruptions occurring during fracturing. The structures observed here are different from those reported in replicas of glutaraldehyde-fixed and glycerol-cryoprotected tissue, in which the intramembrane junctional components partition with the protoplasmic face and often assume the appearance of continuous cylinders. This comparison illustrates some of the artifacts associated with freeze-fracturing and shadowing. On the basis of a comparison of freeze-fracture replicas and sections of lanthanum-infiltrated tissues, the relationship between intramembrane junctional components and intercellular septal elements is analysed.


1996 ◽  
Vol 23 (3) ◽  
pp. 305 ◽  
Author(s):  
MV Sailaja ◽  
VSR Das

Highly characteristic responses of thylakoid membranes were observed in function and composition when fully developed plants of Amaranthus hypochondriacus L. grown under light sufficient (2000 μmol m-2 s-1) conditions were transferred to light limited conditions (650 μmol m-2 s-1 and 200 μmol m-2 s-1). The whole-chain, photosystem I and photosystem II electron transport rates were depressed in both bundle sheath and mesophyll thylakoids with remarkable differences between them in variation of rates under limiting light. The reduction in PSI electron transport in the mesophyll could be attributed to reduced PSI centres, while in the bundle sheath, a modulation of cytochrome b6/f complex regulated the rates of PSI electron transport. The requirement for an unaltered number of PSI centres under limiting light in the bundle sheath is ascribed to operation of an energy-consuming C4 pump.


1970 ◽  
Vol 25 (10) ◽  
pp. 1157-1159 ◽  
Author(s):  
A. Trebst ◽  
E. Harth ◽  
W. Draber

A halogenated benzoquinone has been found to inhibit the photosynthetic electron transport system in isolated chloroplasts. 2·10-6ᴍ of dibromo-thymoquinone inhibit the Hill- reaction with NADP, methylviologen or anthraquinone to 100%, but do not effect the photoreduction of NADP at the expense of an artificial electron donor. The Hill - reaction with ferricyanide is inhibited even at the high concentration of 2·10-5ᴍ of dibromo-thymoquinone to only 60%. The remaining reduction in the presence of the inhibitor reflects the rate of ferricyanide reduction by photosystem II. It is concluded that the inhibition of electron transport by the quinone occurs between photosystem I and II and close to or at the functional site of plastoquinone.


1987 ◽  
Vol 14 (1) ◽  
pp. 21 ◽  
Author(s):  
WS Chow ◽  
AB Hope

The concentrations of photosystem II (PS II), photosystem I (PS I) and cytochrome b/f complexes on a chlorophyll basis have been determined for chloroplasts of spinach grown under three irradiances in a glasshouse. Assaying PS II by atrazine binding gave concentrations that exceeded the estimates from flash-induced O2 or H+ yield by a factor of 1.21 � 0.04 (23). Since part of this factor (1.14) is expected to arise from missed turnovers in excited reaction centres, it is concluded that both methods are valid to determine [PS II] in gently isolated chloroplasts. The agreement between the methods also suggests that atrazine does not bind to additional sites of quenchers such as 'Q400', contrary to the suggestion of R. J. Dennenberg and P. A. Jursinic [Biochim. Biophys. Acta 808 (1985), 192-200]. Whilst [PS I] was independent of growth irradiances, [PS II] and [cyt f ] increased with irradiance, as did the latent ATPase activity, a measure of the concentration of coupling factor 1. If [PS I] is taken as constant at 1.65 mmol (mol Chl)-1, the mean stoichiometries of PS II : cyt b/f complex : PS I obtained at the minimum and maximum irradiances were 1.2 : 0.7 : 1 and 1.6 : 1.0 : 1 respectively, PS II being determined by atrazine binding.


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