Peter J. Quinn

Structural reorganisation of chloroplast thylakoid membranes in response to heat-stress

Abstract Chloroplasts isolated from broad bean (Vicia faba) show major structural reorganisations on heating to temperatures above 35°C. Exposure to increasing temperatures in the range 35–45°;C for 5 min, leads to a progressive destacking of the chloroplast membranes and the replacement of the normal granal arrangement by modified thylakoid attachment sites. An analysis of the size and packing densities of the freeze-fracture particles present in different membrane fracture-faces suggests that this rearrangement reflects the dissociation of the light-harvesting units of Photosystem II. The antennae complexes of Photosystem II appear to cluster together, maintaining regions of membrane adhesion, whilst excluding the core-complexes of Photosystem II and light-harvesting units of Photosystem I from these regions. If the chloroplasts are heated to higher temperatures, 45–55°C, phase-separated aggregates of non-bilayer-forming lipids are often observed. The release of these lipids from their normal constraints within the bilayer is consistent with the idea that they play a role in the packaging of the light-harvesting complexes within the thylakoid membrane.

The structure and thermotropic properties of pure 1,2-diacylgalactosylglycerols in aqueous systems

Pure 3-sn-monogalactosyldilinolenoylglycerol and 3-sn-digalactosyldilinolenoylglycerol have been isolated from bean leaves. Distearoyl derivatives have been prepared by catalytic hydrogenation of the unsaturated galactolipids. The unsaturated lipids form stable monomolecular films at the air/water interface which are similar to liquid-expanded phospholipid monolayers. The limiting areas were about 0.57 nm2 and 0.62 nm2 for the mono- and digalactosyldiacylglycerols, respectively. The saturated galactolipids formed condensed monolayers that were relatively unstable. The surface pressure-area isotherm of the digalactosyl derivative was more expanded than that of the monogalactosyldiacylglycerol especially at low surface pressures. Low-angle X-ray diffraction and freeze-fracture electron microscopy studies of the monogalactosyldiacylglycerols showed that an hexagonal-type structure was formed by the unsaturated lipid in aqueous systems, whilst the saturated lipid was arranged in a lamellar configuration. Both digalactosyldiacylglycerols form lamellar structures in water. A gel-to-liquid-crystalline phase transition of distearoyldigalactosylglycerol was observed at about 51 degrees C by fluorescence depolarization measurements, using 1,6-diphenylhexatriene, and by differential scanning calorimetry. The saturated monogalactosyldiacylglycerol did not form dispersions suitable for fluorescence probe studies of a phase transition. A complex pattern of endotherms was observed for this lipid by differential scanning calorimetry.

The Dynamics Of Membrane Structur

The membranes of living organisms are involved in many aspects of the life, growth and development of all cells. The predominant structural elements of these membranes are lipids and proteins and the basic strucvture of these molecules has been reviewed. The physical properties of the lipid constituents particularly their behavior in aqueous systems has led to the concepts of thermotropic and lyotropic mesomorphism; the interaction between different types of lipid molecules modulate this behavior. Interaction of phospholipids in aqueous systems with cholesterol, ions and drugs have been examined in this context. In addition a variety of model lipid-protein systems have been investigated and the implications of interactions between lipids and different proteins in biological membranes has been evaluated. This leads to a detailed consideration of the way lipids and proteins ae organized in cell membranes and contains an appraisal of the evidence supporting contemporary views of membrane structure. Particular attention has been devoted to the question of how mobile the components are within the structure. Particular attention has been devoted to the question of how mobile the components are within the structure. Finally the biosynthesis, turnover and modulation of the properties of interacting membrane constituents is critically reviewed and possible ways of controlling the behavior of cells and organisms by altering the structural parameters of different membranes has been considered.

The formation of non-bilayer structures in total polar lipid extracts of chloroplast membranes

Abstract The structural organisation of aqueous dispersions of total membrane lipid extracts of broad bean ( Vicia faba ) chloroplasts is dependent on pH and the presence of cations. In the absence of inorganic salts, sonicated dispersions of lipid extract in distilled water form smooth, single-shell vesicles approximately 30–50 nm in diameter. Reducing the pH of the dispersions, to neutralise the acidic lipids present in the extract, or the addition of low concentrations of metal cations, leads to the fusion of the vesicles and a partial phase-separation of the non-bilayer forming lipid monogalactosyldiacylglycerol to form spherical inverted micelles similar to those previously reported for binary mixtures of monogalactosyl and digalactosyldiacylglycerol (Biochim. Biophys. Acta 685, 297–306). Increasing concentrations of polyvalent, but not monovalent, cations lead to further structural rearrangements involving the formation of para-crystalline arrays of tubular and spherical inverted micelles. The factors determining the formation of these different structures, and their possible relevance to the structural organisation of the native chloroplast membrane, are discussed.

Bilayer and non-bilayer transformations in aqueous dispersions of mixed sn-3-galactosyldiacylglycerols isolated from chloroplasts A freeze-fracture study

Abstract A number of different particle and ‘particle-like’ structures are observed in freeze-fracture replicas prepared from aqueous dispersions of mixtures of mono- and digalactosyldiacylglycerol. The smallest of these structures (10–12 nm in diameter) corresponding to inverted lipid micelles sandwiched within lipid bilayers are often organised into extensive planar arrays. A number of larger ‘particle-like’ features are also observed in replicas of this type. An analysis of the relationship between these structures suggests that they reflect responses to stresses associated with a temperature-dependent incorporation of the lipids of the inverted micelles into the lamellar structure.

The polyisoprenoid chain length influences the interaction of ubiquinones with phospholipid bilayers

The interaction between ubiquinone homologues with polyisoprenoid chain lengths varying from 3 to 10 units and dipalmitoylphosphatidylcholine bilayers has been examined by differential scanning calorimetry and wide angle X-ray diffraction analysis. Decreasing the polyisoprenoid chain lengths of ubiquinone in mixed dispersions with phospholipid in mol ratios of about 10 mol% caused a decrease in the gel-liquid crystalline phase transition temperature of the phospholipid and a broadening of the transition. Enthalpy measurements showed that most of the phospholipid (greater than 92%) was involved in the transition endotherm and the formation of a gel phase was also confirmed by the presence of a sharp X-ray reflection of 0.42 nm. These results are consistent with a model in which all of the ubiquinone homologous ultimately undergo a phase separation from phospholipid molecules entering a gel phase on cooling below the phase transition temperature. Reducing the length of the polyisoprenoid chain alters the amphipathic balance of the ubiquinone molecules and is reflected in the tendency of shorter chain ubiquinones to intercalate between the phospholipid molecules upon reheating through the main phase transition.

Phase separations in membranes of Anacystis nidulans grown at different temperatures

Freeze fracture electron microscopy studies were performed on samples of Anacystis nidulans quenched from different temperatures. Membrane lipid phase separations were observed to take place over the ranges 15--30 degrees C, 5--25 degrees C and -5--15 degrees C for cultures grown at 38, 28 and 18 degrees C, respectively. Differential scanning calorimetry heating curves showed endotherms which coincided with these temperature ranges. Variations of phase separation temperatures with growth temperature, and hysteresis effects in the calorimetric measurements, were related to changes in the fatty acid composition of membrane lipids.

The modulation of membrane fluidity by hydrogenation processes. III. The hydrogenation of biomembranes of spinach chloroplasts and a study of the effect of this on photosynthetic electron transport.

A method is reported for the in situ modification of the lipids of isolated spinach chloroplast membranes. The technique is based on a direct hydrogenation of the lipid double bonds in the presence of the catalyst, chlorotris(triphenylphosphine)rhodium (I). The pattern of hydrogenation achieved suggests that the catalyst distributes amongst all of the membranes. The polyunsaturated lipids within the membranes are hydrogenated at a faster rate and at an earlier stage than are the monoenoic lipids. Whilst addition of the catalyst to the chloroplast causes an initial 10--20% decrease in Hill activity, saturation of up to 40% of the double bonds present can be accomplished without causing further significant alterations in photosynthetic electron transport processes or marked morphological changes of the chloroplast structure as observed in the electron microscope.

The modulation of membrane fluidity by hydrogenation processes: II. Homogeneous catalysis and model biomembranes

A homogeneous catalyst, chlorotris (triphenylphosphine) rhodium (I) has been incorporated into model biomembrane structures in the form of lipid bilayer dispersions in water. This enables the hydrogenation of the double bonds of the unsaturated lipids within the bilayers to be accomplished. To decide the optimum conditions for efficient hydrogenation the reaction conditions have been varied. The effect of catalyst concentration, hydrogen gas pressure and lipid composition (with and without cholesterol) have all been studied. The partition of the catalyst into the lipid medium was checked by rhodium analysis. The results show that an increase of catalyst concentration or an increase of hydrogen gas pressure leads to increasing rates of hydrogenation. Successful hydrogenation was accomplished with different types of lipid dispersions (mitochondrial, microsomal and erythrocyte lipids). A selectivity of the homogeneous hydrogenation process is indicated. The polyunsaturated fatty acyl residues are hydrogenated at an earlier stage and at a faster rate than the monoenoic acids. Furthermore, an increase in the proportion of cholesterol to lipid within the bilayer structures causes a progressive decrease in the rate of hydrogenation. The fluidity of the lipid bilayers can be altered to such an extent by the hydrogenation process that new sharp endotherms corresponding to the order-disorder transition of saturated lipids occur at temperatures as high as 319 K. Some potential uses of hydrogenation for the modulation of cell membrane fluidity are discussed as well as the design of new types of catalyst molecules.

The modulation of Ca2+-ATPase activity of sarcoplasmic reticulum by membrane cholesterol. The effect of enzyme coupling

The Ca2+-ATPase activity of sarcoplasmic reticulum is relatively low (less than 2 I.U.) in vesicles where enzyme activity is geared to calcium accumulation. Modulation of membrane fluidity by enriching the membrane with cholesterol has no significant effect on enzyme activity. Collapsing the Ca2+ gradient with the calcium ionophore, A23187, unmasks the inhibitory effect of membrane cholesterol on enzyme activity.