Proton binding to biological membranes.

Biological membranes contain proton-binding moieties. A laser-induced proton pulse was used to characterize the proton-binding properties of bacterioopsin-containing membranes and of sarcoplasmic reticulum. Different protonation and deprotonation processes occurred. The liberation of protons from pyranine dye and the protonation of the membranes were independent of temperature; the reprotonation of pyranine and proton release from the membranes were temperature dependent. In the cases of membrane-free and membrane-containing systems, the activation enthalpies and entropies were calculated from the decay rates. The activation enthalpy of 16 kJ/mol for reprotonation of pyranine in membrane-free solution is characteristic for a diffusion-controlled process. The value for the membrane-containing systems was nearly double, suggesting that the buffering moieties of the membrane surfaces strongly bind the protons, raising the activation enthalpies. This is possibly an effect of the Coulomb cages formed from closely located proton acceptor sites. The activation entropies were positive in all cases.

Photoelectric response of the N intermediate of bacteriorhodopsin and its mutant T46V.

Double flash experiments were performed in order to gain information about the characteristics of the N intermediates of the photocycle of bacteriorhodopsin. The N intermediates of wild-type bacteriorhodopsin and mutant T46V were excited at different delay times after the first laser flash which induced the photocycle and the electric responses were registered. These electric signals revealed that charge motions occurred in both cases, though charge translocation, i.e. H(+) pumping, could not be observed. The delay time dependence of the electric signals is characterized by two distinct processes corresponding to two substates of the N intermediates.

Demonstration of the parity-violating energy difference between enantiomers.

Racemic mixtures of (+) and (-) sodium potassium tartrate, tris(1, 2-ethanediamine)cobalt(III), and tris(1,2-ethanediamine)iridium(III) molecules were crystallized, and the optical activities of the resulting crystalline materials, dissolved in water, were carefully measured to study the influence of the parity-violating energy difference in the crystallization process. Although no effect was found in the case of tartrate, enantiomeric excess appeared in the crystallization of the cobalt and iridium complexes. These investigations, performed in our laboratory, demonstrated the contribution of the parity-violating neutral weak current to the forces acting in molecules.

Morphological and physiological differences between aeroponically and hydroponically grown sunflower plants.

The effects of aeroponic (AP) and hydroponic (HP) conditions on growth rate, morphological traits, potassium uptake and redox activities were compared in sunflower seedlings. Higher growth rate was found under HP than AP conditions and morphological traits were also different. The thickness of AP grown roots increased and new lateral roots with thickened root hairs were formed while the length of AP roots was small. Microscopical studies on cross-sections of the embedded root segments showed that the diameter of cross-section, the diameter of stele and the width of cortex of AP grown roots were significantly higher than those of HP plants. The element composition of AP or HP grown plants also differed, due to the different ion uptake processes. Potassium (86Rb+) uptake of AP grown plants was low and it seemed to be a passive process, while in case of HP grown plants it was decreased by both dinitrophenol and ferricyanide suggesting that the uptake process was coupled to the proton gradient. Roots of AP grown plants showed high ferricyanide reductase activity and it was accompanied by an increased acidification of the medium.

Plasma membrane purification from roots of sunflower by phase partitioning.

Plasma membrane vesicles were purified from the roots of sunflower (Helianthus annuus L. cv. Topflor) by aqueous polymer two-phase partitioning. The optimal conditions for separation were determined by systematic variation of the polymer concentration and salt composition. The phase system containing 6% (w/w) dextran T-500, 6% (w/w) polyethylene glycol 3350, 250 mM sucrose, 5 mM potassium phosphate, pH 7.8, without added salts proved to be the best. The ATPase activity had a pH optimum at 6.5 and it was stimulated by Mg2+, but not by Ca2+. The plasma membrane MgATPase activity was inhibited by vanadate but not by nitrate, an inhibitor of tonoplast ATPase. Only 10% of the microsomal protein was responsible for 36% of the total MgATPase activity. Moreover IDPase activity, a Golgi marker, appeared to be very low indicating the high purity of the preparation.

The effects of detergents on the ATPase activity of plasma membrane prepared by phase partitioning from sunflower roots.

MgATPase activity of plasma membrane from sunflower roots purified by phase partitioning was tested using different permeabilizing agents. MgATPase activity showed similar dependence on Triton X-100, digitonin and Zwittergent 3-14 concentration, the curves resulted from the stimulatory and inhibitory reactions. The latency calculated at their optima was only 52%. In case of n-octyl glucoside the latency was 77%, while the highest latency (82%) was measured in the presence of lysophosphatidylcholine, and it was the only surfactant which did not inhibit the PM ATPase even at 10 times the concentration giving maximum release of latent activity. Therefore, for the further characterization of the PM ATPase lysophosphatidylcholine is suggested to be used as detergent.