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R P Rand

Publications and source records attributed to R P Rand.

At least 91 records · Page 5Linked to original sources
PubMed1982

Interactions between neutral phospholipid bilayer membranes.

We have obtained force vs. separation relations between bilayers in 10 different phospholipid preparations: dilauroyl-dimyristoyl-, dipalmitoyl-, distearoyl-, or dioleoylphosphatidylcholine (PC); egg phosphatidylethanolamine; cholesterol-containing bilayers of dipalmitoyl PC and of egg PC. The chemical potential of water in the multilamellar lattice is determined at all water contents and changes continuously with bilayer separation; no discrete classes of water are observed. The interbilayer van der Waals force is estimated from the balance of forces at the bilayer separation where the multilayer lattice is in equilibrium with pure water. Although quantitative differences are evident for different phospholipids, all force curves but one show a clear, exponentially decaying "hydration "repulsion" whose decay distance is 2-3 A . Estimates of forces between bilayer vesicles show great sensitivity to the identity of the phospholipid polar group and to the packing of the hydrocarbon acyl chains. On major implication of this is the likelihood of local structural changes and lipid segregation in the area of closest approach of interacting vesicles of mixed phospholipids.

Chemical Phenomena↗
PubMed1982

Measurement of the lateral compressibility of several phospholipid bilayers.

Lateral compressibilities of bilayers in multilayer lattices are given for 10 phospholipid preparation:dilauryl-, dimyristoyl-, dipalmitoyl-, distearoyl-, and dioleoylphosphatidylcholine (PC); egg phosphatidylethanolamine (PE); as well as cholesterol-containing bilayers of dipalmitoyl PC or of egg PC. Bilayer deformability is highly nonlinear and does not permit description in terms of a simple modulus. The presence of cholesterol or C=C bonds (dioleoyl PC) increases deformability, but freezing of acyl chains does not cause dramatic stiffening of the bilayer. Lateral compression of dilauryl PC an dimyristoyl PC causes a transition from "melted" to "frozen" acyl chains above the normal transition temperatures. Our measurements do not correspond in any obvious way to lateral compressibilities in monolayers at the air-water interface.

Cholesterol↗
PubMed1981

Agonist potency determination by patch clamp analysis of single glutamate receptors.

Agonists (L-quisqualate, L-glutamate and L-cysteine sulphinate) of the locust muscle glutamate receptor differ in potency. Patch clamp analysis of single glutamate receptors reveals that the relative potencies of agonists are determined both by the life-times of the channels that they gate and their probability of activating the glutamate receptor-ionophore complex. Furthermore, agonists which most readily activate the receptor channel complex gate channels with longer life-times than agonists which are less efficient in this respect.

Animals↗
PubMed1981

Binding of divalent cations of dipalmitoylphosphatidylcholine bilayers and its effect on bilayer interaction.

We have confirmed that CaCl2 swells the multilayer lattice formed by dipalmitolyphosphatidylcholine (DPPC) in an aqueous solution. Specifically, at room temperature 1 mM CaCl2 causes these lipid bilayers to increase their separation, dw, from 19 A in pure water to greater than 90 A. CaCl2 concentrations greater than 4 mM cause less swelling. We have measured the net repulsive force between the bilayers in 30 mM CaCl2 at T = 25 degrees C (below the acyl chain freezing temperature). For interbilayer separations between 30 and 90 A, the dominant repulsion between bilayers is probably electrostatic; Ca2+ binds to DPPc lecithin bilayers, imparting a charge to them. The addition of NaCl to CaCl2 solutions decreases this repulsion. For dw less than 20 A, the bilayer repulsion appears to be dominated by the "hydration forces" observed previously between both neutral and charged phospholipids. From the electrostatic repulsive force, we estimate the extent of Ca2+ binding to the bilayer surface. The desorption and bound Ca2+, apparent when bilayers are pushed together, is more rapid than one would expect if an association constant governed Ca2+ binding. The association affinity does not appear to be a fixed quantity but rather a sensitive function of ionic strength and bilayer separation.

Calcium↗
PubMed1981

Adsorption of divalent cations to a variety of phosphatidylcholine bilayers.

We have determined the degree of binding of divalent cations to several kinds of phosphatidylcholine (PC) bilayers. This has been done by measuring the electrostatic interbilayer repuslive force that results when multilamellar lattices are exposed to Me2+Cl2 solutions. Divalent cations bind to dipalmitoylphosphatidylcholine in the sequence Ca2+ approximately equal to Cd2+ approximately equal to Mn2+ greater than Ca2+ approximately equal to Mg2+ greater than Ba2+. Among the different synthetic lipids, preference for Ca2+ is in the sequence DOPC less than DLPC less than DMPC approximately equal to DPPC approximately equal to DSPC. The density of bound charge is proportional to the density of polar groups on the bilayer surface. Phosphatidylcholines with mixed hydrocarbon chains, such as egg PC or 1:1 mixtures of synthetic PC's, form two distinct lamellar phases in CaCl2 solutions. In all cases the electrostatic force between bilayers decays exponentially with their separation but more slowly than expected from ionic double-layer theory. We suggest that the electric fields from opposing surfaces perturb the zwitterionic charge-binding polar groups and continuously modify their ion binding affinities as the bilayers approach.

Calcium↗
PubMed1980

The effect of cholesterol on measured interaction and compressibility of dipalmitoylphosphatidylcholine bilayers.

We have examined the phase diagram of dipalmitoylphosphatidylcholine (DPPC)--cholesterol-water mixtures at low cholesterol content, and report phase separation between 3 and 10 mol% cholesterol. The two lamellar phases at equilibrium in this region appear to be pure DPPC and 11 mol% cholesterol in DPPC. For these two lamellar phases, which are made up of alternating layers of water and bimolecular lipid leaflets, we have measured the forces of interaction between leaflets and the lateral pressure and compressibility of the leaflets. Both bilayers experience a strong repulsive force when forced together only a few ångströms (1 A = 0.1 nm) closer than their maximum separation in excess water. However, the presence of 11 mol% cholesterol causes the bilayers to move apart of 35-A separation from the 19-A characteristic of pure DPPC in excess water. This swelling may result from a decrease in van der Waals attraction between bilayers or from an increase in bilayer repulsion. Differences in bilayer interaction can be a cause for phase separation. More importantly these differences can cause changes in the composition of regions of membranes approaching contact. At 11 mol%, cholesterol substantially increases the lateral compressibility of DPPC bilayers leading to higher lateral density fluctuations and potentially higher bilayer permeability.

Chemical Phenomena↗
PubMed1979

Measured work of deformation and repulsion of lecithin bilayers.

We used three complementary techniques to vary the chemical potential of water in lipid/water mixtures; we measured the work of removing water from the multilayer lattice formed in water by the zwitterionic phospholipid egg lecithin. By x-ray diffraction, we observed the structural consequences of water removal. There are no discrete classes of "bound water" in this system; the work of removal is a continuous function of water content and lattice repeat spacing. From 30 to 3 A separation between bilayers there exists an exponential "hydration force" repulsion with a 2.6 A decay length. This interaction translates into a very large force to prevent contact between vesicles and planar membranes. It may be an important feature in controlling vesicle-to-cell fusion. As water is removed, bilayers not only move closer, but thicken as the lipid polar groups on the same bilayer move closer together. It is possible to divide the applied work into that of direct bilayer repulsion and that of bilayer deformation. We thus obtained a first determination of the lateral pressure required to create large increases in bilayer thickness and concomitant decreases in bilayer area. The lateral pressure reaches 25 dynes/cm for a 25% decrease in bilayer area. Systematic measurements of the mechanical properties of bilayers suffering such large deformation will allow critical tests of theories on bilayer stability and phase transition.

Mathematics↗
PubMed1978

Measurement of repulsive forces between charged phospholipid bilayers.

By using an osmotic stress technique (LeNeveu, D. M., et al. (1977) Biophys. J. 18, 209), we have measured the net repulsive force between egg lecithin bilayers containing various amounts of the charged lipids phosphatidylglycerol and phosphatidylinositol. At bilayer separations greater than about 30 A, the repulsion is dominated by electrostatic forces; its variation with both bilayer separation and charge density is well described qualitatively by simple electrostatic double-layer theory. Quantitative agreement requires, however, that only about 50% of the phosphatidylglycerol polar groups be dissociated. At all charge densities, even for pure phosphatidylglycerol, and at bilayer separations less than about 30 A, the repulsion is dominated not by the electrostatic force but by a strong "hydration force" (LeNeveu, D. M., et al. (1977) Biophys. J. 18, 209). We conclude that the hydration force demands more attention than it has enjoyed hitherto in attempts to understand bilayer membrane interaction and fusion.

Electrochemistry↗
PubMed1977

Measurement and modification of forces between lecithin bilayers.

We probe in two different ways the competing attractive and repulsive forces that create lamellar arrays of the phospholipid lecithin when in equilibrium with pure water. The first probe involves the addition of low molecular weight solutes, glucose and sucrose, to a system where the phospholipid is immersed in a large excess of water. Small solutes can enter the aqueous region between bilayers. Their effect is first to increase and then to decrease the separation between bilayers as sugar concentration increases. We interpret this waxing and waning of the lattice spacing in terms of the successive weakening and strengthening of the attractive van der Waals forces originally responsible for creation of a stable lattice. The second probe is an "osmotic stress method," in which very high molecular weight neutral polymer is added to the pure water phase but is unable to enter the multilayers. The polymer competes for water with the lamellar lattice, and thereby compresses it. From the resulting spacing (determined by X-ray diffraction) and the directly measured osmotic pressure, we find a force vs. distance curve for compressing the lattice (or, equivalently, the free energy of transfer to bulk water of water between bilayers. This method reveals a very strong, exponentially varying "hydration force" with a decay distance of about 2 A.

Chemical Phenomena↗
PubMed1976

Apparent modification of forces between lecithin bilayers.

Small sugar solutes effect variation in the equilibrium separation of lecithin bilayers in aqueous solution. Since sugars have negligible influence on bilayer structure, they probably act by modifying interbilayer forces. The observed widening and narrowing of the bilayer separation is correlated with the predicted weakening and strengthening of the attractive van der Waals forces between lipid bilayers that occurs with increasing sugar concentrations.

Chemical Phenomena↗
PubMed1976

Perturbation of lecithin bilayer structure by globoside.

The ultrastructure of aggregates formed by mixtures of pig erythrocyte lecithin, cholesterol and globoside in aqueous systems was studied by electron microscopy and X-ray diffraction. Globoside and lecithin in up to equimolar amounts formed a lamellar mesophase, although the structure of the lamellae was perturbed. Mixtures containing excess globoside formed complex tubular or reticular aggregates. Cholesterol appeared to promote mixing of lecithin and globoside. The flexibility gradient of the hydrocarbon (hc) region of the lipid bilayers was studied using electron spin resonance (esr) spectroscopy of various nitroxide-labelled stearic acid probes. Globoside in equimolar amounts greatly perturbed the order parameters of lecithin bilayers, reducing the fluidity of the hc region and flattening the flexibility gradient near the polar (p) surface. The effect of globoside on lecithin-cholesterol bilayers was not so pronounced, since the latter was already more ordered than lecithin bilayers. A phase transition of pure globoside at 55 degrees C, involving 'melting' of the hc chains was also detected using X-ray and esr spectroscopic techniques. The interbilayer spacing, dw, of equimolar lecithin-globoside lamellar phase increased by 42% from that of lecithin bilayers, indicating that the glycolipid p group may increase the net repulsive force between bilayers, as was previously predicted theoretically.

Animals↗
PubMed1975

Tilted hydrocarbon chains of dipalmitoyl lecithin become perpendicular to the bilayer before melting.

Differential scanning calorimetry studies of dipalmitoyl lecithin show two reversible transitions as the temperature is changed between 20 and 50 degrees C. A pretransition endotherm occurs at 35 degrees C prior to the main chain melting endotherm which occurs at 42 degrees C. X-ray diffraction studies show that below 33 degrees C the chains of the lecithin are fully extended, packed in a hexagonal crystalline lattice but tilted with respect to the plane of the bilayer. Between 35 and 42 degrees C the chains are similarly packed but oriented perpendicular to the bilayer plane. Above 44 degrees C the chains are "melted" or disordered. Monolayer studies of dipalmitoyl lecithin using continuous recording of pressure with molecular area reveal the existence of two solid condensed phases corresponding to these tilted and verticle chain structures. The tilted to perpendicular transition would account for the pretransition endotherm of the lipid; the crystalline to melted change corresponds to the larger transition observed at 42 degrees C.

Calorimetry↗
PubMed1975

Lysolecithin and cholesterol interact stoichiometrically forming bimolecular lamellar structures in the presence of excess water, of lysolecithin or cholesterol.

The structural interaction of egg lysolecithin, derived from egg lecithin, and cholesterol in aqueous solution has been investigated using X-ray diffraction. When mixed in any proportions, either suspended in excess buffer or up to 85% lipid by dry weight, a separate lamellar phase containing equimolar proportions of lysolecithin and cholesterol forms, separate from excess water, or lysolecithin or cholesterol. The cholesterol disorders the crystalline chains of the lysolecithin. The equimolar phase is stable up to 50 degrees C unlike lysolecithin alone, which forms micelles, Thes results show that lysolecithin and cholesterol combine stoichiometrically in a stable complex. We propose as a structural model, that cholesterol fills the space of the missing fatty acyl chain making the lysolecithin more cylindrical rather than wedge shaped. This interaction could reduce both the lytic action of lysolecithin on membranes and its induction of cell fusion. It suggest another role of cholesterol in cell membranes: namely, to act as a stabilizer of bilayer structure by being a mobile component that can fill free volume in the hydrocarbon interior. Lysolecithin-cholesterol interaction may also be important in the early events of atherosclerosis where lysolecithin levels in vessel walls increase fivefold.

Binding Sites↗