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PubMed1982

A theory of effect of protons and divalent cations on phase equilibria in charged bilayer membranes: comparison with experiment.

We summarize the concepts in the recently developed statistical mechanical theory of the effects of proton binding and divalent cation binding on phase equilibria in bilayer membrane composed of acidic phospholipids. The theory is used to calculate membrane phase transition temperatures for different aqueous concentrations of protons, divalent cations and monovalent salt. We discuss methods for calculating transition temperatures even for systems in which there is not an excess of protons or divalent cations relative to lipids. The results are compared with existing experimental data for a number of lipids. There is good agreement between calculated transition temperature vs. pH curves and experimental data for dimyristoylmethylphosphatidic acid, dimyristoylphosphatidylglycerol, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidylserine, and dimyristoylphosphatidic acid. General thermodynamic considerations are used to derive in Clapeyron-like equation for the rate of variation in membrane transition temperature with divalent cation concentration. This equation and some available experimental data are used to argue that the large increase in solid to fluid phase transition temperature that is observed experimentally as the divalent cation concentration is increased is the result of the metastable solid phase that exists at low but not high divalent cation concentration. A calculated coexistence diagram is compared with existing experimental data for transition temperatures of dimyristoylphosphatidylglycerol membranes at different total calcium concentrations. Good agreement is obtained when the existence of a metastable solid phase is assumed.

Cations, Divalent↗
PubMed1981

Effect of surface curvature on stability, thermodynamic behavior, and osmotic activity of dipalmitoylphosphatidylcholine single lamellar vesicles.

The size and surface curvature dependence of the properties and stability of single lamellar vesicles have been investigated by using a variety of physicochemical techniques. Dipalmitoylphosphatidylcholine single lamellar vesicles of sizes ranging between 200 and 900 A in diameter have been prepared by the French press method and characterized with respect to their size distribution, stability, and thermotropic behavior by negative stain electron microscopy, molecular sieve chromatography, nuclear magnetic resonance spectroscopy, and differential scanning calorimetry. Vesicles with a diameter smaller than 400 A are unstable below their transition temperature and fuse spontaneously to form larger single lamellar vesicles. Correlation analysis of experimentally obtained size distributions and calorimetric phase transitions profiles allowed estimation of the size dependence of the transition temperature. The phase transition temperature depends on the vesicle size in a sigmoidal fashion. Throughout the entire 200-700 A diamter range, the phase transition parameters are sensitive to size; however, the size dependence is especially pronounced around 400 A in diameter. The anomalous size dependence of the transition temperature for vesicles smaller than 400 A in diameter has been attributed to a decrease in the effective bilayer curvature due to packing rearrangements of the lipid molecules. Changes in the fractional degree of self-quenching of trapped 6-carboxyfluorescein induced by osmotic stress indicate that large single lamellar vesicles are not spherical under isoosmotic conditions. These vesicles are relatively flexible and can sustain almost a 2-fold increase in their internal aqueous volume without any leakage of the internal content.

Drug Stability↗
PubMed1990

Interaction of cyclosporin A with dipalmitoylphosphatidylcholine.

Cyclosporin A, a hydrophobic cyclic peptide, is a potent immunosuppressant. In an attempt to determine the localization of cyclosporin A in phospholipid membranes, the effect of cyclosporin A on dipalmitoylphosphatidylcholine (DPPC) has been investigated using deuterium nuclear magnetic resonance (2H-NMR) spectroscopy and differential scanning calorimetry (DSC). Cyclosporin A was dispersed within acyl chain per-deuterated DPPC at a concentration of 6 mole percent, hydrated with buffer, and the spectra obtained over a range of temperatures were compared with that of pure DPPC. The changes caused by cyclosporin A were assessed by the first moment (M1) and order parameters calculated from the spectra. The presence of cyclosporin A decreases the magnitude of M1 at temperatures below the gel to liquid-crystalline phase transition temperature but increases M1 at temperatures above the transition. In addition, the change in M1 at the transition temperature was also less abrupt when cyclosporin A was present. For bilayers in the liquid-crystalline state, cyclosporin A causes an increase in the order parameters along the acyl chains which suggests that cyclosporin A is located along the acyl chains of the phospholipid. For DSC, cyclosporin A was dispersed in non-deuterated DPPC at different peptide to phospholipid mole ratios. The endothermic peaks associated with the gel to liquid-crystalline phase transition and pretransition were recorded and compared with similar mole ratios of cholesterol to lipid. At 30 mole percent cyclosporin A, small decreases in the main transition temperature and associated enthalpy were observed, whereas at 30 mole percent cholesterol, the main transition is barely distinguishable from the baseline. The pretransition was not observed with the addition of 11 mole percent of either cyclosporin A or cholesterol. The results of the thermal analysis indicate that although cyclosporin A and cholesterol appear to be both located along the acyl chains of the phospholipids, they have dramatically different interactions with the membrane lipids.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed2005

Determination of pore sizes and volumes of porous materials by 129Xe NMR of xenon gas dissolved in a medium.

In our previous paper (J. Phys. Chem. B 2005, 109, 757) it was illustrated that the 129Xe NMR spectra of xenon dissolved in acetonitrile confined into mesoporous materials give detailed information on the system, especially about the pore sizes. A resonance signal originating from xenon atoms sited in very small cavities built up inside the pores during the freezing transition (referred to as signal D) turned out to be highly sensitive to the pore size. The emergence of this signal reveals the phase transition temperature of acetonitrile inside the pores, which can also be used to determine the size of the pores. In addition, the difference in the chemical shifts of two other signals arising from xenon dissolved in bulk and confined acetonitrile (B and C) provides another method for determining the pore sizes. In the present work, the observed correlations have been investigated using an extensive set of measurements with a variety of porous materials (silica gels and controlled pore glasses) with the mean pore diameters ranging from 43 to 2917 A. The usefulness of the correlations has been demonstrated by calculating the pore size distributions from the spectral data. The distributions are in agreement with those reported by the manufacturers, when the mean pore diameter is smaller than approximately 500 A. In addition, it has been shown that the porosity of the materials can be determined by comparing the intensities of the signals arising from the bulk and confined liquid. When acetonitrile is replaced by cyclohexane in the sample, the dependence of the chemical shift difference between the B and C signals on the pore size becomes more sensitive, but no D signal appears below the freezing point. In addition, the influence of xenon gas on the melting points of bulk and confined acetonitrile has been studied by 1H NMR cryoporometry. The measurements show that the temperature of the latter transition lowers slightly more, and consequently affects the pore sizes calculated by means of the difference in the phase transition temperatures. Hysteresis in the phase transitions in a cooling-warming cycle has also been studied as a function of the temperature stabilization time by 129Xe NMR of xenon dissolved in acetonitrile.

Journal Article↗
PubMed1976

Spin-labeling studies on the lipids of psychrophilic, psychrotrophic, and mesophilic clostridia.

Spin-labeling studies were conducted to elucidate the viscosity and phase transition temperatures of lipids isolated from psychrophilic, psychrotrophic, and mesophilic clostridia. Electron spin resonance spectroscopy indicated that the lipids, for all the growth temperatures tested, were in a fluid state and from 13 to 24 C higher than the corresponding lipid transition temperatures. When the organisms were grown at different temperatures, a psychrotropic and two mesophilic clostridia were shown to be able to adjust their lipid-phase transition temperature to the growth temperature. A psychrophilic Clostridium strain, when grown at different temperatures, synthesized lipids that had the same phase transition temperature. It is suggested that this lack of growth temperature-inducible regulation of lipid-phase transition temperature may be a molecular determinant for the psychrophily of this organism. It is proposed that the growth temperature range of an organism is dependent upon the ability of the organism to regulate its lipid fluidity within a specific range.

Clostridium↗
PubMed2004

Thermally activated nanowear modes of a polymer surface induced by a heated tip.

We report on nanoscale wear induced by atomic force microscopy using a heated cantilever/tip on a 20 nm thick film of polystyrene. Wear modes dependent on tip temperature characteristics were identified. Below the glass transition temperature of the polymer, the formation of quasi one-dimensional surface ripples with a typical period on the order of 100 nm was observed. We found that the ripple height typically saturates at 20 nm and that the buildup rate depends on temperature and load. From these observations we can calculate an activation energy for the ripple formation, which is on the order of 0.4 eV, at temperatures close to but below the glass transition temperature of the polymer. In the glass transition regime the ripple formation is strongly enhanced. An abrupt change of the wear mode is observed as the polymer in contact with the tip is heated above the glass transition temperature. Here, polymer material is transported along with the propagating tip.

Journal Article↗
PubMed1997

Membrane fatty acid composition and membrane fluidity as parameters of stress tolerance in yeast.

The relationship among membrane fatty acid composition, membrane fluidity, and stress tolerance was investigated in yeast cells. Several strains were examined for their ability to survive heat, ethanol, and hydrogen peroxide stresses. Membrane fluidity was determined by measuring fluorescence anisotropy using diphenylhexatriene as a probe. There was no obvious relationship among membrane fatty acyl composition, membrane fluidity, and stress tolerance in the strains examined. A consistent trend in the present study was an observed decrease in membrane fluidity following thermal treatment, which coincided with a reduction in cell viability. We suggest that protein denaturation may be responsible for the observed effect of elevated temperature on membrane fluidity and viability. This was implied by observations on the irreversible nature of thermal transitions, as measured by breaks in Arrhenius plots, in which stationary phase cells were shown to exhibit higher transition temperatures (53.9-55.5 degrees C) than exponential phase cells (49.5-51 degrees C). Furthermore, the thermal transition temperature was shown to increase in exponential phase cells following heat shock, which was associated with an increase in thermotolerance. We suggest that the thermotolerant state of heat-shocked cells and cells entering stationary phase may be associated with increased protein stability. However, despite the relatively good correlation between thermal transition temperature and stress tolerance, the thermal transition temperature did not predict the stress tolerance of a given strain, as stress-sensitive strains had similar transition temperatures to those of stress-resistant strains.

Cell Membrane↗
PubMedDate not supplied

Ethanol and temperature effects on five membrane bound enzymes.

The effects of ethanol on the activities of five membrane bound enzymes were determined using a crude membrane preparation obtained from cortex of long-sleep (LS) and short-sleep (SS) mice. These two mouse lines were selectively bred for differences in duration of ethanol-induced sleep time. The enzymes studied were two forms of NaK-ATPase, Mg-ATPase, 5'nucleotidase, and acetylcholinesterase. Arrhenius plots of the ethanol-temperature-enzyme activity studies indicate specificity in ethanol's actions. NaK-ATPase activity consists of two enzymes which were distinguished by sensitivity to ouabain. The Arrhenius plot of the high ouabain sensitivity enzyme (low Ki) exhibited a transition temperature which was reduced twice as much by ethanol in LS membranes as in SS membranes. Ethanol did not affect the transition temperature of the high Ki NaK-ATPase but the control (no ethanol) transition temperature was 2.7 degrees higher in SS membranes. Arrhenius plots of Mg-ATPase activity did not exhibit a transition temperature and ethanol did not alter enzyme activity. Ethanol did not alter the transition temperatures of 5'nucleotidase or acetylcholinesterase but the control transition temperature for acetylcholinesterase was 2.3 degrees higher in SS membranes. These results indicate specificity in ethanol's actions on membranes and that inhibition of the lipid-enzyme interactions for the low Ki NaK-ATPase is correlated with the difference in sensitivity to ethanol seen between the LS and SS mice.

5'-Nucleotidase↗
PubMed1994

Differential scanning calorimetry of thermotropic phase transitions in vitaminylated lipids: aqueous dispersions of N-biotinyl phosphatidylethanolamines.

The thermotropic phase behavior of a homologous series of saturated diacyl phosphatidylethanolamines in which the headgroup is N-derivatized with biotin has been investigated by differential scanning calorimetry. In 1 M NaCl, derivatives with acyl chainlengths from C(12:0) to C(20:0) all exhibit sharp chain-melting phase transitions, which are reversible with a hysteresis of 1.5 degrees or less, except for the C(12:0) lipid which has a transition temperature below 0 degree C. The transition enthalpy and the transition entropy depend approximately linearly on the lipid chainlength, with incremental values per CH2 group that are very similar to those obtained for the corresponding underivatized phosphatidylethanolamines in aqueous dispersion. The chainlength-independent contribution to the transition enthalpy is significantly smaller than that for the underivatized phosphatidylethanolamines, and that for the transition entropy is much smaller; the latter suggesting that the N-biotinylated phosphatidylethanolamine headgroups are differently hydrated from those of the underivatized lipids. The gel-to-fluid phase transition temperatures of the N-biotinylated lipids are lower than those of the parent phosphatidylethanolamines, and their chainlength dependence conforms well with that predicted by assuming that the transition enthalpy and entropy are linearly dependent on chainlength. Although the chain-melting phase behavior is generally similar to that of the parent phosphatidylethanolamines, the gel phases (and the fluid phases in the case of chainlengths C(12:0) to C(16:0)) have a different lyotropic structure in the two cases, and this is reflected in the chainlength-independent contributions to the thermodynamic parameters. In the absence of salt, the thermotropic phase behavior of aqueous dispersions of the N-biotinyl phosphatidylethanolamines is considerably more complex. The transition temperatures are consistently lower than those in 1 M NaCI, but the transitions are broader, contain multiple peaks and exhibit a much larger hysteresis between heating and cooling scans. Additionally, the lipids with shorterchainlengths exhibit metastability in the absence of salt, converting from a micellar solution to a lamellar gel phase only after incubation at low temperature with freeze-thaw cycling.

Biotin↗
PubMed1979

Phase properties of senescing plant membranes: role of the neutral lipids.

Wide-angle X-ray diffraction studies have indicated that rough and smooth microsomal membranes from bean cotyledons acquire increasing proportions of gel phase lipid at physiological temperature as the tissue senesces. In addition, for both types of membrane the lipid phase transition temperature, defined as the highest temperature at which gel phase lipid can be detected, progressively rises with advancing senescence. Liposomes prepared from total lipid extracts of the membranes show a similar increase in transition temperature with age, indicating that separation of the polar lipids into distinct gel and liquid-crystalline domains is not attributable to peculiar protein-lipid interactions. Liposomes prepared from purified phospholipid fractions of the membranes show little change in transition temperature with age, indicating that the altered phase properties of the lipid do not reflect an increase in fatty acid saturation. However, the formation of gel phase lipid that occurs naturally during senescence can be stimulated by preparing liposomes from a mixture of the phospholipid fraction from young membrane and the neutral lipid fraction from old membrane. By adding the separated components of the neutral lipid fraction to purified phospholipid it was found that sterol esters and several unidentified lipids are able to raise the transition temperature of the polar lipids. Sterols have no effect on the phospholipid transition temperature. The data have been interpreted as indicating that several neutral lipids, which presumably increase in abundance with advancing senescence, induce a lateral phase separation of the polar lipids resulting in distinct gel and liquid-crystalline domains of lipid in the senescent membranes.

Fabaceae↗
PubMed1987

Effects of halothane on dipalmitoylphosphatidylcholine liposomes: a Raman spectroscopic study.

Raman spectroscopy has been used to monitor the concentration of halothane (1-bromo-1-chloro-2,2,2-trifluoroethane) in 20% aqueous dispersions of dipalmitoylphosphatidylcholine (DPPC) as well as to follow changes in the acyl chain order within the hydrocarbon interior of the liposomes. Temperature profiles for the gel to liquid-crystalline phase transitions for the liposomes were constructed from changes in peak height intensity ratios in the C-H stretching mode and C-C stretching mode regions. Halothane present at the clinical level produces a change of -0.5 degrees C in the phase transition temperature. A limiting transition temperature of about 21 degrees C and saturation of the gel phase occur when the molar ratio of halothane to DPPC reaches about 1.25. At molar ratios above 2.1, the liquid-crystalline phase is also saturated with halothane. Calculations of the distribution of halothane between the various phases in the system are presented and used to interpret literature data as well as the present experiments. Ideal solution theory accounts rather well for the depression in the transition temperature over most of the mole ratio range, an outcome which implies that halothane is excluded from the hydrocarbon interior but not the head-group region in the gel phase. The role of halothane in the head-group region is discussed.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1980

Sodium-potassium-activated adenosine triphosphatase activity as a measure of neuronal membrane characteristics in ethanol-tolerant mice.

(Na+-K+) activated adenosine triphosphatase of mouse synaptosomal membranes is inhibited by high concentrations of ethanol. When membranes were obtained from mice made tolerant to and physically dependent on ethanol by chronic exposure to an ethanol-containing liquid diet, the enzyme was resistant to the inhibitory effects of ethanol. Arrhenius plots of synaptosomal (Na+-K+) activated adenosine triphosphatase from control animals revealed that ethanol added in vitro lowered the transition temperature and altered the Arrhenius activation energies of this enzyme. Enzyme from ethanol-tolerant animals had a lower transition temperature than that from control animals, and ethanol added in vitro had no effect either on transition temperature or on activation energy of enzyme from ethanol-tolerant animals. The occurrence of lowered transition temperature and resistance to ethanol-induced alterations in transition temperature of the enzyme from ethanol-tolerant animals most likely reflects changes in membrane composition. Changes in Arrhenius plots correlated in time with resistance to the inhibitory effects of ethanol on (Na+-K+) activated adenosine triphosphatase activity and the time course of disappearance of these effects was similar to that of the disappearance of functional tolerance to ethanol. The resistance to the effects of ethanol on membrane function may be related to ethanol tolerance evidenced by behavioral and physiological measurements.

Alcoholism↗
PubMed1978

Studies on the effect of the lipid phase transition on the interaction of glucagon with dimyristoyl glycerophosphocholine.

Glucagon is found to interact with dimyristoyl glycerophosphocholine both above and below the phase transition temperature of the lipid. Above the phase transition temperature the interaction is manifested by an increase in the rate of vesicle aggregation and by an increased permeability of unilamellar vesicles to Eu3+ and to Fe(CN)3-6. However, no stable lipoprotein complex can be detected by gel filtration. Below the phase transition glucagon can form stable complexes with dimyristoyl glycerophosphocholine vesicles but cannot rapidly rearrange these vesicles to disk-shaped particles until the phase transition temperature is approached. The energy of activation for the dissociation of glucagon from the disk-shaped lipoprotein particle is 29 kcal/mol at temperatures above 36 degrees C but increases markedly at lower temperatures, as the region of the lipid phase transition is approached. This increase in energy of activation at lower temperatures is most probably due to the larger amount of energy required to rearrange gel-state lipid in the transition state and provides an explanation for the unusual kinetic stability of the glucagon-dimyristoyl glycerophosphocholine lipoprotein complex only at temperatures below the phase transition of the lipid.

Chemical Phenomena↗
PubMed2003

Difference and similarity of dielectric relaxation processes among polyols.

Complex permittivity measurements were performed on sorbitol, xylitol, and sorbitol-xylitol mixture in the supercooled liquid state in an extremely wide frequency range from 10 microHz to 500 MHz at temperatures near and above the glass transition temperature. We determined detailed behavior of the relaxation parameters such as relaxation frequency and broadening against temperature not only for the alpha process but also for the beta process above the glass transition temperature, to the best of our knowledge, for the first time. Since supercooled liquids are in the quasi-equilibrium state, the behavior of all the relaxation parameters for the beta process can be compared among the polyols as well as those for the alpha process. The relaxation frequencies of the alpha processes follow the Vogel-Fulcher-Tammann manner and the loci in the Arrhenius diagram are different corresponding to the difference of the glass transition temperatures. On the other hand, the relaxation frequencies of the beta processes, which are often called as the Johari-Goldstein processes, follow the Arrhenius-type temperature dependence. The relaxation parameters for the beta process are quite similar among the polyols at temperatures below the alphabeta merging temperature, T(M). However, they show anomalous behavior near T(M), which depends on the molecular size of materials. These results suggest that the origin of the beta process is essentially the same among the polyols.

Journal Article↗
PubMed1998

Effect of single chain lipids on phospholipase C-promoted vesicle fusion. A test for the stalk hypothesis of membrane fusion.

The effect of low proportions (up to 5 mol %) of single-chain lipids on phospholipase C-promoted fusion of large unilamellar vesicles has been investigated with the aim of testing the so-called stalk model of membrane fusion. This model is known in two main versions, the one originally published by Kozlov and Markin [Kozlov, M. M. and Markin, V. S. (1983) Biofizika 28, 255-261] and what is known as the "modified stalk model" [Siegel, D. P. (1993) Biophys. J. 65, 2124-2140], that differ in a number of predictions. In the view of the latter author, hydrocarbons or other nonpolar lipids should help fusion by decreasing the interstitial energy of the stalk connecting the two apposed bilayers. We show that small amounts of hexadecane or squalene increase significantly the fusion rates in our system. Changes in monolayer curvature are the object of different predictions by the original and modified stalk theories. According to the original form, fusion would be promoted by lipids inducing a negative curvature in the closest (cis) monolayers of the fusing membranes and inhibited by the same lipids in the trans monolayers; the opposite would happen with lipids inducing a positive curvature. The modified stalk model predicts that fusion is helped by increasing the negative curvature of both monolayers. In our system, symmetrically distributed arachidonic acid, which increases the negative curvature, enhances lipid and content mixing, and the opposite is found with symmetrically distributed lysophosphatidylcholine or palmitoylcarnitine, which facilitate a positive monolayer curvature. In addition, fluorescence polarization and 31P NMR studies of the lamellar-to-isotropic (Q224 cubic) thermotropic transition of a lipid mixture corresponding to our liposomal composition reveal that all lipids that facilitate fusion decrease the transition temperature, while fusion inhibitors increase the transition temperature. Moreover, fusion (content mixing) rates show a maximum at the lamellar-to-isotropic transition temperature. These observations support the involvement of inverted lipid structures, as occurring in the inverted cubic phases, in membrane fusion. All these data are in full agreement with the stalk model of membrane fusion, particularly in its modified version.

Alkanes↗
PubMedDate not supplied

Thermoresponsive drug delivery using liquid crystal-embedded cellulose nitrate membranes.

The aim of this study was to investigate the use of thermotropic liquid crystalline (TLC) blends of 4-pentyl-4'-cyanobiphenyl (K15) and 4-heptyl-4'-cyanobiphenyl (K21) with appropriate nematic to isotropic phase temperature (Tn - i) just above body temperature as a temperature-modulated drug permeation system. Using differential scanning calorimetry (DSC) we showed that the phase transition temperature (Tn - i) of K15 and K21 were 34.2 degrees C and 41.5 degrees C respectively. However, the thermogram of K15 and K21 blends with different ratios was shown to be a single endothermic peak similar to that of pure TLCs. K15 and K21 blends did not behave as a physical blend of two thermotropic liquid crystals with different Tn - i. However, they are rather mixed together in such ways that behave like a single unit TLC. The Tn - i of these TLC mixtures was linearly proportionate to the ratio of K15:K21. Using appropriate ratio of K15:K21 TLC, a mixture with desirable phase transition temperature was obtained. A triple layer of cellulose nitrate membranes containing a 50:50 mixture of K15 and K21 was used for drug permeation studies. This composite membrane showed good pulsatile permeation of drug molecules in response to temperature changes below and above the Tn - i of the K15 and K21 blends in a reproducible and reversible manner. Paracetamol and methimazole were chosen as hydrophobic and hydrophilic drug models, respectively. Methimazole permeability through the TLC membrane was much higher (36.0 x 10(-5) cm/s) at temperatures above the phase transition temperature of liquid crystal blends than that (7.2 x 10(-5) cm/s) at temperatures below the phase transition temperature of liquid crystal blends (38.1 degrees C).

Acetaminophen↗
PubMed2002

The origin of the anomalous superconducting properties of MgB(2).

Magnesium diboride differs from ordinary metallic superconductors in several important ways, including the failure of conventional models to predict accurately its unusually high transition temperature, the effects of isotope substitution on the critical transition temperature, and its anomalous specific heat. A detailed examination of the energy associated with the formation of charge-carrying pairs, referred to as the 'superconducting energy gap', should clarify why MgB(2) is different. Some early experimental studies have indicated that MgB(2) has multiple gaps, but past theoretical studies have not explained from first principles the origin of these gaps and their effects. Here we report an ab initio calculation of the superconducting gaps in MgB(2) and their effects on measurable quantities. An important feature is that the electronic states dominated by orbitals in the boron plane couple strongly to specific phonon modes, making pair formation favourable. This explains the high transition temperature, the anomalous structure in the specific heat, and the existence of multiple gaps in this material. Our analysis suggests comparable or higher transition temperatures may result in layered materials based on B, C and N with partially filled planar orbitals.

Journal Article↗
PubMed1985

Interaction of the sperm adhesive protein, bindin, with phospholipid vesicles. I. Specific association of bindin with gel-phase phospholipid vesicles.

Bindin is a 30,000-mol-wt protein of sea urchin sperm that is responsible for the specific adhesion of the sperm acrosomal process to the vitelline layer covering the egg plasma membrane during fertilization. Sulfated glycoconjugates are believed to be the egg surface receptors for bindin, but the mechanism by which bindin associates with the sperm acrosomal membrane is unknown. Here I report that bindin specifically associates with phospholipid vesicles in vitro. Interaction of the bindin polypeptide with liposomes was found to cause an increase in the density of the liposomes and induce the aggregation of the vesicles. A novel property of this association of bindin with membranes was that it required phospholipids in a gel phase. The interaction of bindin with liposomes was greatly reduced at temperatures above the phase transition temperature. The interaction of bindin with gel-phase vesicles appeared to be reversible, since the aggregated vesicles dissaggregated as the temperature was raised above the phase transition temperature. Association of bindin with the bilayer did not alter the accessibility of the polypeptide to cleavage by trypsin, which suggests that most of the polypeptide chain remains exposed at the surface of the membrane.

Animals↗