DOPG

A clear physical picture of the dynamic behavior of molecules on the surface of the lipid membrane is highly desired and has attracted great attention from researchers. In this study, a step forward in this direction based on previous studies was presented with second harmonic generation (SHG) and molecular dynamic (MD) simulation. Specifically, details on the orientation flipping and cross-membrane transport of two charged molecules, 4-(4-diethylaminostyry)-1-methyl-pyridinium iodide (D289) and malachite green (MG), on the surface of 2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG) lipids were presented. Firstly, the orientation flipping of the two molecules on the surface of lipids before their cross-membrane transport was confirmed by the MD simulation. Then, the concentration dependent rate of the cross membrane transport for MG/D289 was analyzed. It was found that a simplified model could satisfactorily interpret the faster cross-membrane transport of MG under higher bulk concentrations. A different concentration dependent dynamics was observed with D289 and the reason behind it was also discussed. With this investigation, the surface structures and dynamics of D289 and MG on the DOPG lipid surface were clearly presented.

In cells, the breakdown of arginine to ornithine and ammonium ion plus carbon dioxide is coupled to the generation of metabolic energy in the form of ATP. The arginine breakdown pathway is minimally composed of arginine deiminase, ornithine transcarbamoylase, carbamate kinase, and an arginine/ornithine antiporter; ammonia and carbon dioxide most likely diffuse passively across the membrane. The genes for the enzymes and transporter have been cloned and expressed, and the proteins have been purified from Lactococcus lactis IL1403 and incorporated into lipid vesicles for sustained production of ATP. Here, we study the kinetic parameters and biochemical properties of the individual enzymes and the antiporter, and we determine how the physicochemical conditions, effector composition, and effector concentration affect the enzymes. We report the KMand VMAXvalues for catalysis and the native oligomeric state of all proteins, and we measured the effect of pathway intermediates, pH, temperature, freeze-thaw cycles, and salts on the activity of the cytosolic enzymes. We also present data on the protein-to-lipid ratio and lipid composition dependence of the antiporter.

Irreversible electroporation (IRE) is a technique for the disruption of localized cells or vesicles by a series of short and high-frequency electric pulses which has been used for tissue ablation and treatment in certain diseases. It is well reported that IRE induces lateral tension in the membranes of giant unilamellar vesicles (GUVs). The GUVs are prepared by a mixture of anionic lipid dioleoylphosphatidylglycerol (DOPG) and neutral lipid dioleoylphosphatidylcholine (DOPC) using the natural swelling method. Here the influence of DOPG mole fraction, XDOPG, on the critical tension of electroporation in GUVs has been investigated in sodium chloride-containing PIPES buffer. The critical tension decreases from 9.0 ± 0.3 to 6.0 ± 0.2 mN/m with the increase of XDOPGfrom 0.0 to 0.60 in the membranes of GUVs. Hence an increase in XDOPGgreatly decreases the mechanical stability of membranes. We develop a theoretical equation that fits the XDOPGdependent normalized critical tension, and obtain a binding constant for the lipid-ion interaction of 0.75 M-1. The decrease in the energy barrier for formation of the nano-size nascent or prepore state, due to the increase in XDOPG, is the main factor explaining the decrease in critical tension of electroporation in vesicles.

The cell plate is the new cell wall, with bordering plasma membrane, that is formed between two daughter cells in plants, and it is formed by fusion of vesicles (approximately 60 nm). To start to determine physical properties of cell plate forming vesicles for their transport through the phragmoplast, and fusion with each other, we microinjected fluorescent synthetic lipid vesicles that were made of 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DOPG) into Tradescantia virginiana stamen hair cells. During interphase, the 60-nm wide DOPG vesicles moved inside the cytoplasm comparably to organelles. During cytokinesis, they were transported through the phragmoplast and accumulated in the cell plate region together with the endogenous vesicles, even inside the central cell plate region. Because at this stage microtubules are virtually absent from that region, while actin filaments are present, actin filaments may have a role in the transport of vesicles toward the cell plate. Unlike the endogenous vesicles, the synthetic DOPG vesicles did not fuse with the developing cell plate. Instead, they redistributed into the cytoplasm of the daughter cells upon completion of cytokinesis. Because the redistribution of the vesicles occurs when actin filaments disappear from the phragmoplast, actin filaments may be involved in keeping the vesicles inside the developing cell plate region.