Small Head Group Research Articles

The Anionic Phospholipid-Mediated Membrane Interaction of the Anti-Cancer Drug Doxorubicin Is Enhanced by Phosphatidylethanolamine Compared to Other Zwitterionic Phospholipids

The interaction of doxorubicin and lipids has been studied using large unilamellar vesicles (LUVET) composed of mixtures of anionic phospholipids and various zwitterionic phospholipids. Dilution of anionic lipids with zwitterionic lipids leads to decreased membrane association of the drug because electrostatic forces are very important in doxorubicin-membrane interaction. However, binding of doxorubicin to LUVET composed of anionic phospholipids combined with phosphatidylethanolamine (PE) is much higher than binding to LUVET made of anionic lipids plus a range of other zwitterionic lipids such as phosphatidylcholine (PC) and the N-methylethanolamine and N, N-dimethylethanolamine derivatives of PE. This preferential interaction is observed with all negatively charged phospholipids tested and is, in the case of phosphatidylserine (PS), confirmed in monolayer experiments. The increase in surface area observed in a monolayer composed of PS and PE (1/3) was 3 times higher than in a monolayer of PS/PC (1/3). The preferential interaction appears not to be due to the ability of PE to adopt inverted nonbilayer structures, but probably involves a combination of the ability of PE to form additional hydrogen bonds and of the intrinsic curvature of a bilayer containing PE because of its small headgroup. Implications of our finding for the in vivo membrane interaction and transport of the drug will be discussed.

Packing constraints and electrostatic surface potentials determine transmembrane asymmetry of phosphatidylethanol

The energetic determinants of the distribution of anionic phospholipids across a phosphatidylcholine (PtdCho) bilayer with different packing constraints in the two leaflets were studied, using (13)CH2-ethyl-labeled phosphatidylethanol (PtdEth) as a (13)C NMR membrane probe. PtdEth is unique in exhibiting a split (13)CH2-ethyl resonance in sonicated vesicles, the two components originating from the inner and outer leaflets, thus permitting the determination of the PtdEth concentration in each leaflet. Small and large unilamellar PtdEth-PtdCho vesicles were prepared in solutions of different ionic strengths. A quantitative expression for the transbilayer distribution of PtdEth, based on the balance between steric and electrostatic factors, was derived. The transbilayer difference in packing constraints was obtained from the magnitude of the PtdEth signal splitting. The electrostatic contribution could be satisfactorily described by the transmembrane difference in Gouy-Chapman surface potentials. At low (0.1-0.25%) PtdEth levels and high (up to 500 mM) salt concentrations, PtdEth had a marked fivefold preference for the inner leaflet, presumably because of its small headgroup, which favors tighter packing. At higher PtdEth content (4.8-9.1%) and low salt concentrations, where electrostatic repulsion becomes a dominant factor, the asymmetry was markedly reduced and an almost even distribution across the bilayer was obtained. In less curved, large vesicles, where packing constraints in the two leaflets are approximately the same, the PtdEth distribution was almost symmetrical. This study is the first quantitative analysis of the balance between steric and electrostatic factors that determines the equilibrium transbilayer distribution of charged membrane constituents.

Influence of the head group size on the direction of tilt in Langmuir monolayers

A model of rods with heads of variable size, which are confined to a planar surface, is used to study the influence of the head group size on tilted phases in Langmuir monolayers. Simple free energy considerations as well as exact zero temperature calculations indicate that molecules with small head groups tilt towards next nearest neighbors, and molecules with larger head groups towards nearest neighbors. This provides a possible explanation for recent experimental results, and for details of the generic phase diagram for fatty acid monolayers.

Critical dependence of the solubilization of lipid vesicles by the detergent CHAPS on the lipid composition. Functional reconstitution of the nicotinic acetylcholine receptor into preformed vesicles above the critical micellization concentration

The critical concentration of free detergent [D w] c, which is necessary for lipid vesicle solubilization, is widely believed to be near to the critical micellization concentration (CMC) of the detergent. Here it is shown that [D w] c and the critical detergent/lipid ratio R c(M) in the mixed micelles strongly depend on the lipid composition. In agreement with the concept of packing constraints, phospholipids with a large head-group were solubilized by the detergent CHAPS at low CHAPS concentrations, for example [D w] c = 0.27 mM for phosphatidyl-inositol and [D w] c = 2.3 mM for phosphatidylcholine, T = 293 K (20°C). In contrast, phospholipids (PL) with a small head-group required larger [CHAPS] values for (mixed) micelle formation: [D w] c = 3.2 mM for phosphatic acid (PA) and [D w] c = 5.2 mM for a mixture of palmitoyloleoylphosphatidylcholine (POPC), phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), POPC:PE:PG = 30:60:10 (mol.-%). Values obtained for the partition coefficient K = R c (M) [ D w] c ranged from K = 0.12 mM −1 for dimyristoylphosphatidylcholine to K = 1.1 mM −1 for (soy) phosphatidylcholine:phosphatidylglycerol (50:50). After addition of 30 mol.-% cholesterol (Ch) or with dipalmitoylphosphatidylcholine below T = 308 K (35°C), [D w] c is > 10 mM, which is far above the CMC = 4 mM. This indicates that CHAPS micelles are formed before vesicle solubilization begins. The analysis of vesicle solubilization was a prerequisite for the controlled reconstitution of protein membrane proteins. AChR proteins reconstituted into preformed PL Ch -vesicles at [D w] > CMC had a 4–5 time higher value of Li-influx compared to reconstitutions from completely solubilized lipid and proteins indicating a higher efficiency of right-side out AChR incorporation.

Brij 58, a polyoxyethylene acyl ether, creates membrane vesicles of uniform sidedness. A new tool to obtain inside‐out (cytoplasmic side‐out) plasma membrane vesicles

Most of the plasma membrane vesicles formed upon homogenization of plant tissue have a right-side-out (cytoplasmic side-in) orientation. Subsequent purification of plasma membrane vesicles using aqueous two-phase partitioning leads to a further enrichment in right-side-out vesicles resulting in preparations with 80-90% of the vesicles in this orientation. Thus, to be able to assay, e.g. the ion-pumping activities of the H(+)-ATPase and the Ca(2+)-ATPase, which expose their active sites towards the cytoplasm, the vesicles have to be inverted. This is very efficiently achieved by including 0.05% of the detergent Brij 58 (C16E20) in the assay medium, which produces 100% sealed, inside-out (cytoplasmic side-out) vesicles from preparations of 80-90% right-side-out vesicles. This was shown by assaying ATP-dependent H+ pumping using the delta pH probe acridine orange and dissipating the H+ gradient with nigericin, and by assaying ATP-dependent Ca2+ transport using 45Ca2+ and dissipating the Ca2+ gradient with the ionophore A23187. The presence of intact vesicles was confirmed by electronmicroscopy. The detergent Brij 58 is a polyoxyethylene acyl ether and a survey among some other members of this series revealed that those with a head group of relatively large size (E20-23) showed this 'non-detergent behavior', whereas those with smaller head groups (E8-10) behaved as normal detergents and permeabilized the membranes. Thus, a very convenient system for studies on ion-pumping activities and other vectorial properties of the plasma membrane is obtained by simply including the detergent Brij 58 in the assay medium.

Modulation of rhodopsin function by properties of the membrane bilayer

A prevalent model for the function of rhodopsin centers on the metarhodopsin I (MI) to metarhodopsin II (MII) conformational transition as the triggering event for the visual process. Flash photolysis techniques enable one to determine the [MII]/[MI] ratio for rhodopsin in various recombinant membranes, and thus investigate the roles of the phospholipid head groups and the lipid acyl chains systematically. The results obtained to date clearly show that the p K for the acid-base MI-MII equilibrium of rhodopsin is modulated by the lipid environment. In bilayers of phosphatidylcholines the MI-MII equilibrium is shifted to the left; whereas in the native rod outer segment membranes it is shifted to the right, i.e., at neutral pH near physiological temperature. The lipid mixtures sufficient to yield full photochemical function of rhodopsin include a native-like head group composition, viz, comprising phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS), in combination with polyunsaturated docosahexaenoic acid (DHA; 22:6ω3) chains. Yet such a native-like lipid mixture is not necessary for the MI-MII conformational transition of rhodopsin; one can substitute other lipid compositions having similar properties. The MI-MII transition is favored by relatively small head groups which produce a condensed bilayer surface, viz, a comparatively small interfacial area as in the case of PE, together with bulky acyl chains such as DHA which prefer a relatively large cross sectional area. The resulting force imbalance across the layer gives rise to a curvature elastic stress of the lipid/water interface, such that the lipid mixtures yielding native-like behavior form reverse hexagonal ( H II) phases at slightly higher temperatures. A relatively unstable membrane is needed; lipids tending to form the lamellar phase do not support full native-like photochemical function of rhodopsin. Thus chemically specific properties of the various lipids are not required, but rather average or material properties of the entire assembly, which may involve the curvature free energy of the membrane-lipid water interface. These findings reveal that the membrane lipid bilayer has a direct influence on the energetics of the conformational states of rhodopsin in visual excitation.

Peculiarities in the aggregative behaviour of gangliosides, glucosidic surfactants of biological origin

Glucosidic surfactants play an important role in living matter. An interesting class of them are the gangliosides which have a double-tail hydrophobic part, like phospholipids, and a ramified oligosaccharide chain as polar head. The large hydrophobic moiety ensures very low CMC values, of the order of 10−8 M. Phospholipids have a small head group compared with their hydrophobic part. Therefore they form mainly bilayer-type structures in water. Gangliosides, instead, may form either micelles or bilayers depending on the length and conformation of the oligosaccharide chain. For long chains, micelles are formed with a large aggregation number and a long lifetime (many hours), while vesicles are found for short sugar chains. The presence of a large head group makes vesicle mechanical properties rather peculiar. For instance, the ganglioside GM3, which has the longest sugar chain compatible with bilayer-type structures, forms vesicles spontaneously in solution without supply of external energy. This means that the bending rigidity of the GM3 bilayer is very low, allowing very large thermal undulations. GM3 vesicles, made of a single surfactant, are then found to be in thermodynamic equilibrium with a small amount of large bilayer and multilamellar structures. Frustration of the single-component vesicle system is released if a second amphiphile, of the same type of the ganglioside GM3 but with a larger head group, is added. In this case, spontaneous-curvature readjustements of the two monolayers via demixing can give rise to a spontaneous curvature of the bilayer, energetically favouring vesicles over other structures. It is, in fact, observed experimentally that by adding increasing proportions of GM1 the large aggregates gradually diminish in number, until a pure vesicle solution is obtained for a GM1 to GM3 ratio of 35 to 65.

Nonionic Surfactant Vesicles: A Study of Vesicle Formation, Characterization, and Stability

The aggregation state of monoalkyl polyoxyethylene ether surfactants was investigated. This study mainly focused on the aggregation of surfactant molecules into vesicular structures, which can be used as drug delivery systems. The physicochemical parameters that play a role in vesicle formation were determined by varying both the hydrophobic and the hydrophilic moiety of the amphiphilic molecules systematically. A wide range of these nonionic surfactants was studied. The alkyl chain length was varied between 10 and 18 carbon atoms, and the length of the polyoxyethylene head group was varied from 3 to 7 ethyleneoxide units. It appeared that stable vesicle suspensions could only be obtained using surfactants that form liquid state bilayers, and contain a relatively small head group compared to the alkyl chain. These surfactants exhibit a lamellar phase in the water rich regions of the water/surfactant phase diagram. If the head group were too large, micelles instead of vesicles were formed. However, after addition of 40 mol% cholesterol vesicle formation was possible for all surfactants used. It appeared that the size of the vesicles is dependent on the preparation method, but can also be controlled by shaking the suspension vigorously. The nonionic surfactant vesicles were stable at room temperature for a period of at least two months. Electrolytes such as NaCl and phosphate appeared to have an effect on the bilayer stacking of the vesicles.

Interfacial tension studies on surfactant systems at the aqueous/perfluorocarbon interface

The effect of different phospholipid based emulsifier systems on dynamic interfacial tension at the perfluorocarbon/aqueous interface was investigated using a polytetrafluoroethylene Wilhelmy plate method. A standard phospholipid mixture composed of synthetic analogs of the major molecular species present in lecithin was formulated to represent natural egg yolk lecithin in these studies. Natural egg yolk lecithin and Pluronic F68 were studied for comparison with the standard phospholipid mixture. Analogs of the minor phospholipid species present in lecithin and other surfactant systems were investigated as additives to the standard phospholipid mixture as follows: phosphatidylinositol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, cholesterol, cholesterol oleate and Pluronic F68. The perfluorocarbon/distilled water interface is a non-interacting system and therefore ideal for use in interfacial characterization of the different phospholipids and investigation of the effect of slight changes in composition on the interfacial properties of phospholipid mixtures. Low concentrations of the surfactant systems were used so that small changes in the interfacial properties could be detected on addition of the various additives. The equilibrium interfacial tension value of the standard phospholipid mixture increased on the addition of charged phospholipids; phospholipids with short fatty acyl groups and phospholipids with small head groups. The addition of Pluronic F68, lysophosphatidylcholine, lysophosphatidylethanolamine, cholesterol and cholesterol oleate to the standard phospholipid mixture resulted in lower dynamic interfacial tension values.

Retention Mechanisms of Self-Associating Compounds on Sephadex Columns

The monomers of heterocyclic compounds and surfactants having a small head group and a long alkyl chain are adsorbed on the Sephadex gels G-10 and G-200. The adsorption of the former compounds is striking, whereas that of the latter compounds is weak and decreases with increasing CMC.

The transit sequence mediates the specific interaction of the precursor of ferredoxin with chloroplast envelope membrane lipids.

The interaction of the precursor of the chloroplast protein ferredoxin with membrane lipids was studied in monolayer experiments in order to investigate the possible involvement of membrane lipids in the protein translocation process. The precursor efficiently and specifically inserts into a total lipid extract of its biological target the outer envelope membrane of chloroplasts. This interaction is mediated by the transit sequence as it can also be observed for the chemically prepared transit peptide of ferredoxin but neither for the ferredoxin apoprotein nor holoprotein. Interactions with the individual chloroplast lipids, monogalactosyl-diacylglycerol, sulfoquinovosyl-diacylglycerol, and phosphatidylglycerol are predominantly involved which corresponds to the results obtained for transit peptide fragments of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (van't Hof, R., Demel, R. A., Keegstra, K., and De Kruijff, B. (1991) FEBS Lett. 291, 350-354). No efficient interaction was obtained with digalactosyl-diacylglycerol and phosphatidylcholine, suggesting that a loose lipid headgroup packing due to small lipid headgroups and/or electrostatic repulsions facilitates efficient insertion. The observed preferences for interaction of the precursor and transit peptide of ferredoxin for the chloroplast outer envelope membrane lipid extract and the presequence of cytochrome c oxidase subunit IV for the mitochondrial outer membrane lipid extract indicate that targeting sequence-lipid interactions contribute to organelle-specific protein targeting.

A molecular dynamics study of the packing structures in monolayers of partially fluorinated amphiphiles

We report the results of molecular dynamics simulations of liquid-supported monolayers of three partially fluorinated amphiphile molecules, namely CF3(CF2)9CH2COOH, CF3(CF2)6CH2(CF2)3COOH, and CF3(CF2)6(CH2)4COOH. These studies were undertaken to provide information on the interplay between molecular flexibility and the packing structure in a monolayer so as to better interpret the results of recent experiments. The qualitative aspects of the predictions of the simulations are consistent with the recent experimental data for monolayers of CF3(CF2)9CH2COOH [S. W. Barton, A. Goudot, O. Boulassa, F. Rondelez, B. Lin, F. Novak, A. Acero, and S. A. Rice, J. Chem. Phys. 96, xxx (1992)]. In particular, the observed breakup of the homogeneous ordered monolayer into ordered islands with the same collective tilt of the molecules is correctly predicted, and the fact that the collective tilt angle is small is correctly predicted. However, the experimental and theoretical values of the tilt angles are not in quantitative agreement, which we attribute to the inadequacy of the atom–atom potentials used in the simulations. In general, for monolayers of CF3(CF2)9CH2COOH we find that the collective tilt angle predicted is a sensitive function of the area per molecule and is smaller than in monolayers of alkane alcohols and alkane acids. The results of the simulations of monolayers of other partially fluorinated species suggest that the difference in size between the fluorocarbon segments and the smaller head groups or flexible ‘‘spacer’’ CH2 segments can generate subtle changes in the packing structure of a monolayer and the relative stabilities of the untilted and tilted structures.

Evidence for spontaneous segregation phenomena in mixed micelles of gangliosides

A light scattering study of the effect of mixing in aqueous solution two gangliosides, GM2 and GT1b, having different hydrophilic headgroups and similar lipid moieties is presented. Mixed micelle formation with spatial segregation of one ganglioside with respect to the other was observed. It is also shown that segregation is a spontaneous phenomenon which is explainable only in terms of simple geometrical arguments, that is by the fact that the large headgroup of GT1b provides the lipidic core of the aggregate with a better shielding from water in the highly curved regions than the smaller headgroup of GM2 can do. This finding may be of help in understanding the behaviour of gangliosides in artificial and natural membranes.

Equilibrium and dynamic bilayer structural properties of unsaturated acyl chain phosphatidylcholine-cholesterol-rhodopsin recombinant vesicles and rod outer segment disk membranes as determined from higher order analysis of fluorescence anisotropy decay.

Limited-frequency phase-modulation fluorometry of diphenylhexatriene (DPH) and trimethylammonium-diphenylhexatriene (TMA-DPH) was used to characterize the equilibrium and dynamic lipid structural properties of (1) reconstituted egg phosphatidylcholine (egg PC)-rhodopsin vesicles varying in rhodopsin content from 0 to approximately 1 mol %, (2) reconstituted PC-cholesterol-rhodopsin vesicles containing approximately 1 mol % rhodopsin and 0, approximately 15, or approximately 30 mol % cholesterol with egg PC, DOPC (di-18:1-PC), or PAPC (16:0,20:4-PC) as the phospholipid constituent, and (3) native bovine rod outer segment disk membranes. Experiments were conducted at 37, 25, 15, and 5 degrees C. Fluorescence lifetime analysis was performed by fitting the data to a constrained, discrete, biexponential model. Rotational depolarization properties were considered by a model requiring a single rotational diffusion coefficient and capable of producing orthogonal, bimodal orientational distributions for DPH and unimodal distributions for TMA-DPH [Straume, M., & Litman, B. J. (1987) Biochemistry 26, 5113-5120]. Unbleached rhodopsin reduced mean fluorophore lifetimes in proportion to the amount of protein present in PC vesicles as a result of probe-to-retinal energy transfer by (1) redistributing the relative lifetime contributions in favor of the short lifetime population and (2) reducing the lifetimes of each derived population. Lifetimes were increased by cholesterol and by reduction of the temperature, but the relative proportions of derived short- and long-lifetime populations were not affected. TMA-DPH lifetimes were more sensitive (in a relative manner) than were those of DPH. These observations are interpreted in terms of cholesterol and reduced temperature each inhibiting water penetrability into these bilayers, with a greater effect occurring in the headgroup and interfacial regions (probed by TMA-DPH) than in the hydrophobic bilayer interior (probed by DPH). Diunsaturated DOPC-rhodopsin recombinants were more resistant to temperature-dependent lifetime changes than were mixed-chain egg PC or PAPC vesicles. This suggests less favorable interaction of rhodopsin with diunsaturated PCs than with mixed-chain PCs. Lifetimes in disk membranes exhibited this same temperature dependence although DPH in disks had lifetimes longer than those seen in recombinant vesicles. TMA-DPH lifetimes in disks were more similar to those observed in cholesterol-containing recombinants. It would therefore appear that the large proportion of small, charged (at pH 7) phosphatidylethanolamine and phosphatidylserine headgroups present in disks reduces water penetrability into the d

Influence of the alcohol and surfactant structure on the extent of the monophasic domains for some water/oil/alcohol/surfactant systems

The influence of the alcohol and surfactant structure on the extent of the monophasic domain of pseudoternary phase diagrams of water/oil/cosurfactant/surfactant has been investigated. It is shown that surfactants with about 12–16 carbon atoms in the alkyl chain and small head group size are the most suitable for the formation of microemulsions. It is also shown that the combination of 1-butanol with toluene and 1-pentanol with dodecane gives the largest extent of the monophasic domain.

Hydration of ionic surfactant micelles from water oxygen-17 magnetic relaxation

Water oxygen-17 relaxation rates have been measured for aqueous solutions of micelles composed of ionic surfactants of varying alkyl chain length, head group, and counterion. The concentration of surfactant and salt was also varied. From these measurements, supplemented with relaxation data for short-chain molecules and with quadrupolar splittings from anisotropic mesophases, we derive structural and dynamic information about the molecular details of the water-micelle interaction. Water molecules at the micelle surface reorient anisotropically, typically 2-3 times slower than in pure water. The average lifetime for water molecules associated with sodium dodecyl sulfate micelles is 6-37 ns. The water-hydrocarbon contact in the micellar solutions is equivalent to less than two fully exposed methylene groups per amphiphile. Small head groups and small counterions produce the largest effects on the 17O relaxation rate, as expected from geometrical and electrostatic considerations.

Characterization of micellar and liposomal dispersions of gangliosides and phospholipids.

Gangliosides are minor surface components of most mammalian cells, where they are located mainly in the outer leaflet of the lipid bilayer of the plasma membrane (1,2, 3). They are also present in membranes of some enveloped viruses (4). In membranes they serve as receptors for various toxins, viruses, hormones and their pattern is often drastically changed in neoplasia (1, 3, 5). The gangliosides, which are part of the lipid bilayer of the membranes differ in their lyotropic behaviour from the phospholipids and cholesterol which constitute the main lipid components of the membrane (6,38). The membrane phospholipids have two long hydrophobic chains, an interface region and a relatively small inogenic head group. They are classified as “non-soluble swelling amphipaths (6), implying that they do not form micelles but disperse spontaneously in water, forming bilayered multilamellar large liposomes (MLV) or, upon ultrasonic irradiation small unilamellar vesicles (SUV). In contrast, the gangliosides are “soluble amphipaths” (6) which form micelles in water (7). The fact that, in spite of their two long hydrophobic chains the gangliosides are classified as “soluble” amphipaths is explained by their large and highly negatively charged polar head group. Since gangliosides and phospholipids differ in their state of aggregation, their coexistance in membranes may affect or even disturb the bilayered structure. This study aimed to investigate the mutual relations between gangliosides arrl the main lipid components of the membranes. For this purpose the structural and dynamic properties of dispersions composed of well-characterized membrane lipids (either synthetic or of natural sources) and well defined, pure gangliosides were studied with the aid of physical and enzymatic methods.

Aggregation of amphiphiles as micelles or vesicles in aqueous media

The physical factors responsible for the aggregation of amphiphiles in aqueous media are examined and expressions for their contribution to the attractive or repulsive components of the free energy change of aggregation are established. Whereas in previous treatments, an arbitrary repulsive force was necessary to explain the behavior of nonionic systems, no such ad hoc assumption is made here. Rather the free energy changes due to interfacial tension at the hydrocarbon core (of the aggregates)-water interface and to the loss of a part of translational and rotational degrees of freedom of the amphiphiles when they aggregate are the two main repulsive contributions. On the other hand, the factors favoring aggregation are: (i) the van der Waals interactions between the hydrocarbon tails of the amphiphiles, and (ii) the structural changes in water and the changes in the interactions between amphiphiles and water resulting from aggregation. For ionic and zwitterionic amphiphilar systems additional free energy contributions are included to account for the repulsive electrostatic interactions between the head groups. For vesicles, the repulsion caused by the overlapping electrical double layers inside the vesicles is also considered. The expressions established for the various free energy changes associated with aggregation are used to examine the formation of micelles and vesicles, from single and double chain amphiphiles with nonionic, ionic, or zwitterionic head groups. In general, single chain amphiphiles aggregate as micelles, rather than as vesicles, for all types of polar head groups. Depending upon the nature of the head groups small and/or large micelles can form. Nonionic amphiphiles which have head groups of small cross-sectional areas form large micelles, whereas those with large cross-section aggregate as small micelles. This happens because the repulsion caused by the loss of translational degrees of freedom is larger in the latter of the two cases. Ionic or zwitterionic amphiphiles form small micelles even though they have small head groups because of the electrostatic repulsion between the head groups. At large ionic strengths, large micelles can form because the repulsive interactions between the head groups are small. Nonionic double chain amphiphiles aggregate predominantly as vesicles. Ionic or zwitterionic double chain amphiphiles aggregate as micelles when the electrostatic repulsion between the head groups is large and as vesicles when this repulsion is small. However for intermediate values of these interactions both micelles and vesicles form depending upon the length of the hydrocarbon tail. Most biologically significant double chain amphiphiles have long, complex polar head groups and they aggregate as micelles when the hydrocarbon tail length is short, even if the electrostatic repulsion between the head groups is weak; but they aggregate as vesicles when the hydrocarbon tail length is long. The size distributions calculated for different types of amphiphiles can be unimodal, representing a single population of aggregates, bimodal, or trimodal representing the coexistence of two or three distinct populations of aggregates. The three possible populations are small micelles, large micelles, and vesicles.

Enzymology of Ubiquinone‐Utilizing Electron Transfer Complexes in Nonionic Detergent

The enzymology of isolated succinate: ubiquinone reductase and ubiquinone: cytochrome c reductase in nonionic detergents (alkyl polyoxyethylene derivatives) was studied. In the membrane the two multiprotein complexes and their hydrophobic substrates ubiquinone and dihydroubiquinone, are embedded in a common lipid bilayer. In detergent solutions the complexes are each inserted into micelles. Detergent micelles also serve as a solvent for the complexes hydrophobic substrates. As a consequence the isolated complexes are in a discontinuous phase with respect to their hydrophobic substrates and with respect to each other. Three types of assays were used. Firstly, single enzyme assays in which the hydrophobic substrates had to transfer from free micelles to the complex-bound micelles in order for enzyme reactions to occur. Secondly, assays in which the enzymic reactions were coupled to auxiliary nonenzymic reactions which rapidly converted the hydrophobic products back into substrates within the complex-bound micelle. Dichloroindophenol was used for the oxidation of dihydroubiquinone and dihydroduroquinone for the reduction of ubiquinone. Thirdly, assays in which the succinate: ubiquinone reductase reaction was coupled with the ubiquinone: cytochrome c reductase reaction. With the first type of assay, the kinetics of the substrate transfer reaction was dependent upon the type of detergent. In detergents with small polyoxyethylene head groups the transfer reactions were rate-limiting, and in detergents with large polyoxyethylene head groups the transfer reactions were fast and the enzymic reactions were rate-limiting...

Theory of lipid monolayer and bilayer phase transitions: effect of headgroup interactions.

Headgroup and soft core interactions are added to a lipid monolayer-bilayer model and the surface pressure-area phase diagrams are calculated. The results show that quite small headgroup interactions can have biologically significant effects on the transition temperature and the phase diagram. In particular, the difference in transition temperatures of lecithins and phosphatidyl ethanolamines is easy to reproduce in the model. The phosphatidic acid systems seem to require weak transient hydrogen bonding which is also conjectured to play a role in most of the lipid systems. By a simple surface free energy argument it is shown that monolayers under a surface pressure of 50 dynes/cm should behave as bilayers, in agreement with experiment. Although the headgroup interactions are biologically very significant, in fundamental studies of the main phase transition in lipids they are secondary in importance to the hydrocarbon chain interactions (including the excluded volume interaction, the rotational isomerism, and the attractive van der Waals interaction).