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M C Blok

Publications and source records attributed to M C Blok.

12 recordsLinked to original sources

Association of bacteriorhodopsin with lipid-impregnated filters. Evidence for fusion of bacteriorhodopsin-containing vesicles with the lipid phase of the filter.

Bacteriorhodopsin vesicles were associated with cellulose-nitrate filters impregnated with a solution of phospholipids in hexadecane. The generation of (photo)potentials upon illumination of the filter was studied in the absence and presence of ionophores, phospholipase A2, EDTA or polyene antibiotics. From these experiments the following conclusions are drawn. 1. Upon illumination of the filter, bacteriorhodopsin pumps protons into aqueous compartments located in the filter. 2. These aqueous compartments possibly do not originate from the compartments enclosed by the bacteriorhodopsin vesicles. Evidence is obtained that aqueous compartments are present in the surface layers of the lipid-impregnated filters. 3. The results are explained most easily by a mechanism, whereby fusion occurs between the vesicles and the lipids of the filter.

Amphotericin B

Light-induced pH changes inside bacteriorhodopsin vesicles as measured by 31 P NMR.

31P NMR has been used to measure light-induced pH changes inside bacteriorhodopsin vesicles containing entrapped sodium glucose-6-phosphate. Reversible light-induced pH changes were observed at various pH values. The results indicate that our vesicle preparations were not homogeneous with respect to the generation of pH gradients.

Bacteriorhodopsins

A molecular basis for an irreversible thermodynamic description on non-electrolyte permeation through lipid bilayers.

The non-electrolyte permeability of liposomal membranes has been investigated according to the concepts of irreversible thermodynamics. A strong interaction between the permeation of solute and water was observed. This solute-solvent interaction can be fully described by assuming that a number of water molecules will copermeate with each molecule of solute. This number of copermeating water molecules is independent of the nature of the permeant and of temperature, but depends on the osmotic concentration of impermeants inside the liposomes.

Glucose

Non-electrolyte permeability as a tool for studying membrane fluidity.

1. The reflection coefficient for the permeation of thiourea through bilayers of phosphatidylcholine is a function of the fatty-acid composition of the lipid molecules. By means of these reflection coefficients an index for membrane fluidity has been given to each of those lipids, relative to that of egg phosphatidylcholine. 2. The maximum number of water molecules that can copermeate with each molecule of solute by means of solute-solvent interaction is a function of the packing of the lipid molecules in the bilayer. This parameter has been used in this paper for characterizing the fluidity of cholesterol-containing membranes and for membranes with their lipids in the gel state.

Chemical Phenomena

The effect of cholesterol incorporation on the temperature dependence of water permeation through liposomal membranes prepared from phosphatidylcholines.

The permeation of water through liposomal membranes composed of phosphatidylcholine plus varying amounts of cholesterol was studied as a function of temperature. 1. Increasing amounts of cholesterol caused a gradual disappearance of the abrupt change in the rate of water permeation near the gel to liquid-crystalline phase transition temperature of dipalmitoylphosphatidylcholine and dimyristoylphosphatidylcholine liposomes. At cholesterol concentrations above about 30 mol % there was no longer a discontinuity in the rate of water permeation. 2. The incorporation of cholesterol produces a steep change in the activation energy of the water permeation above the transition temperature of the saturated lecithin occurring at about 15 mol % of cholesterol. Below the transition temperature there was a gradual decrease in the activation energy of the water permeation in the region of 0 to 33 mol % of cholesterol. 3. In systems containing unsaturated phosphatidylcholines cholesterol also enhanced the activation energy of the water permeation although to a lesser extent. The results indicate that the position of the cis-double bond in the fatty acid chain is very important in this respect. 4. In systems in which cholesterol increased the temperature dependence of the water permeation there is also an enhancement of the temperature dependence of the isotonic glycerol and erythritol swelling by the same number of kcal/mol.

Biological Transport

Effect of the gel to liquid crystalline phase transition on the osmotic behaviour of phosphatidylcholine liposomes.

Aspects of osmotic properties of liposomes, prepared from synthetic lecithin, above, at and below the gel to liquid crystalline phase transition temperature are described. The experiments show that liposomal membranes with their lipids in the gel state are still permeable to water. The rate of water permeation changes drastically on passing the transition temperature. The water permeation has activation energies of 9.5 +/- 1.28 and 26.4 +/- 0.9 kcal/mol above and below the transition temperature, respectively, indicating that the diffusion processes take place by different mechanisms. With respect to the barrier properties of the liposomes in the vicinity of the transition temperature, the following conclusions can be made. (1) Studying the osmotic shrinkage of liposomes at a fixed temperature near the transition point, the experiments indicate that dimyristoyl phosphatidylcholine membranes are highly permeable to glucose under these conditions, where liquid and solid domains co-exist. Under the same conditions the osmotic experiments did not indicate a strong increase in glucose permeability of dipalmitoyl phosphatidylcholine membranes as compared to the situation above and below the transition temperature. (2) On the other hand, perturbations of the phase equilibrium by temperature varations resulted in a marked increase of the glucose permeation through dipalmitoyl phosphatidylcholine bilayers. Once a new phase equilibrium of liquid and solid regions is established the permeation rate of glucose is much less.

Binding Sites

The effect of chain length and lipid phase transitions on the selective permeability properties of liposomes.

This paper describes experiments showing the importance of the fatty acid chain length on the barrier properties of liposomal bilayers, prepared from saturated lecithins, under conditions of lateral phase separation. 1. Above the gel to liquid crystalline phase transition temperature, liposomes prepared from saturated lecithins with 14 or more carbon atoms per acyl chain exist as stable bilayers, which are practically impermeable to ions. 2. At temperatures well above the transition temperature dilauroyl phosphatidylcholine liposomes exhibited osmotic shrinkage, which was dependent on the ionic size of the solute used to bring about the osmotic gradient, indicating that the permeation through these less stable bilayers takes place mainly via individual diffusion of the permeating ions. 3. An enhanced release of trapped potassium from liposomes was demonstrated in the vicinity of the transition temperature. The extent of the increase, however, depended strongly on the length of the paraffin chain. 4. From measurements of the shrinkage behaviour of liposomes in the vicinity of the transition temperature it is concluded that the increased permeability decreases with increasing diameter of the permeating ion. This finding implies that the increased permeability at the transition temperature cannot be ascribed to "macroscopic" rupture of the liposomal membrane. The maximum permeability in the vicinity of the Tc is discussed in terms of probability and size distribution of statistical pore formation at the boundaries of liquid and solid domains.

Cations, Monovalent