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Results for “Phase Separation”

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At least 217 records · Page 12Linked to original sources

PubMed2002

Pattern formation in phase-separating gels with spontaneous shear.

We study pattern formation in gels undergoing simultaneous phase separation and orientational ordering. A 2D numerical simulation is performed using a minimal model of nonlinear elasticity with density-anisotropy coupling. For strong positive coupling, the collapsed phase elongates along the phase boundary and buckle, creating a folded structure with paired topological defects. For negative coupling, soft elasticity of the swollen phase causes a droplet morphology as in liquid-liquid phase separation. Their possible realizations in nematic liquid-crystalline gels are discussed.

Journal Article↗

PubMedDate not supplied

Isolation of human erythrocyte acetylcholinesterase using phase separation with Triton X-114 and monoclonal immunosorbent chromatography.

A generally applicable approach to the preparative isolation of amphiphilic membrane proteins that follow the Triton X-114 phase during a temperature-dependent phase separation is described. The phase separations were performed direct on whole blood and a 650-fold purification of human erythrocyte membrane acetylcholinesterase (AchE) was obtained. Thus, 0.2 mg enzyme was isolated per 1 liter of blood, with a specific activity of 13 IU/mg, the major contaminants being glycophorin and hemoglobin. The protein material was isolated from the detergent phase by Cu2+ chelate chromatography. This material was used to raise monoclonal anti-AchE antibodies which, when applied to immunosorbent chromatography of washed Triton X-100-lysed erythrocytes in one step, allowed a 246,000-fold purification of AchE with a yield of 88% and a specific activity of 3800 IU/mg.

Acetylcholinesterase↗

PubMed1983

Polymyxin B-induced phase separation and acyl chain interdigitation in phosphatidylcholine/phosphatidylglycerol mixtures.

Monolayers, fluorescence polarization, differential scanning calorimetry and X-ray diffraction experiments have been carried out to examine the effect of the polypeptide antibiotic polymyxin B on the phase behaviour of dipalmitoylphosphatidylglycerol (DPPG) either pure or mixed with dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC). It is shown that in both phosphatidylglycerol alone and phosphatidylglycerol/phosphatidylcholine mixtures, polymyxin B can induce either phase separation between lipid domains of various compositions or interdigitation of the acyl chains in the solid state, without segregation of the two lipids. Phase separation was observed by fluorescence and differential scanning calorimetry after addition of the antibiotic to vesicles composed of mixtures of DMPC and DPPG in conditions where polymyxin B did not saturate phosphatidylglycerol (DPPG to polymyxin B molar ratio, Ri, higher than 15). Phase separation was also observed in mixed monolayers of DPPC and of the 5:1 DPPG/polymyxin B complex, at high surface pressure. Acyl chain interdigitation was observed by X-ray diffraction in both 5:1 DPPG/polymyxin B mixtures and preformed 5:5:1 DMPC/DPPG/polymyxin B mixture, in which the antibiotic saturates phosphatidylglycerol (Ri 5). In both cases, raising the temperature gave rise to a complex double-peaked phase transition by differential scanning calorimetry, from the interdigitating phase to a normal L alpha lamellar phase. As it is known that polymyxin B does not interact with phosphatidylcholine, the data presented show that, when phosphatidylcholine and phosphatidylglycerol are mixed together, a phase perturbation such as acyl chain interdigitation, which normally affects only phosphatidylglycerol, is also felt by phosphatidylcholine.

Calorimetry, Differential Scanning↗

PubMed2006

Effect of the cluster size on the micro phase separation in mixtures of beta-lactoglobulin clusters and kappa-carrageenan.

The phase separation of globular protein clusters formed by heat-denatured beta-lactoglobulin (beta-lg) in mixtures with the polysaccharide kappa-carrageenan (kappa-car) has been studied at pH 7 and 20 degrees C. The effect of the protein cluster size on the phase separation was investigated by preparing clusters with radii between 20 nm and 1 mum. The formation of protein rich microdomains led to an increase of the turbidity starting at a minimum kappa-car concentration that decreased with increasing cluster size, but was only weakly dependent on the protein concentration. The size and number of microdomains do not depend much on the cluster size, but their density decreases with increasing cluster size leading to a lower turbidity.

Carrageenan↗

PubMed2003

Phase separation in polyfluorene-polymethylmethacrylate blends studied using UV near-field microscopy.

In this paper we present a near-field microscopy study of thin films of a phase-separated blend of the fluorescent conjugated-polymer poly(9,9-dioctylfluorene) [PFO] with the non-fluorescent polymer polymethylmethacrylate [PMMA]. A scanning near-field optical microscope (NSOM) was used to generate (blue) fluorescence from the PFO following UV excitation at 362 nm. A range of different concentrations of PFO in PMMA were studied ranging from 1 to 50% PFO in PMMA by mass. By studying both the shear force and fluorescence images we were able accurately to determine the distribution of PFO in the PMMA. We found that phase separation occurs over a number of different length-scales between 5 micro m and 250 nm. We show that at PFO concentrations of 1%, the PFO lies on top of the PMMA. At a PFO relative concentration of 50%, the PMMA phase extends through the whole thickness of the film to the underlying substrate. We use such samples to discuss the resolution of NSOM when imaging thick organic films. Furthermore, we confirm that the length-scales of phase separation can be modified via control over spin-casting protocols.

Equipment Design↗

PubMed2005

Electrochemical detection of nanoscale phase separation in binary self-assembled monolayers.

Developing methods to probe the nature and structure of nanoscale environments continues to be a challenge in nanoscience. We report a cyclic voltammetry investigation of the internal, hydrogen-bond-driven phase separation of amide-containing thiols and alkanethiols. Amide-containing thiols with a terminal ferrocene carboxylate functional group were investigated in two binary monolayers, one homogeneously mixed and the other phase separated. The electrochemical response of the ferrocene probe was used to monitor adsorbate coverage, environment, and phase separation within each of these monolayers. The results demonstrate that the behavior of ferrocene-containing monolayers can be used to probe nanoscale organization.

Journal Article↗

PubMed1993

Role of lateral phase separation in the modulation of phospholipase A2 activity.

Phospholipase A2-catalyzed hydrolysis of phosphatidylcholine large unilamellar vesicles is characterized by a period of slow hydrolysis followed by a rapid increase in the rate of hydrolysis. The temporal relationship between the burst of PLA2 activity and the lateral distribution of substrate and product lipids was examined by simultaneously recording product accumulation and the fluorescence of 1-pyrenyldecanoate, a fatty acid derivative sensitive to lipid distribution and lateral diffusion. The excimer: monomer ratio of the probe changes slowly prior to the burst in activity and then abruptly at the time of the burst. A partial phase diagram for the ternary codispersion of substrate and products (dipalmitoylphosphatidylcholine and 1:1 monopalmitoylphosphatidylcholine/palmitic acid) was constructed by differential scanning calorimetry and suggests gel/gel immiscibility in this system. Thus, the changes in pyrene fluorescence during the time course of hydrolysis appear to be due to lateral phase separation. The critical mole fraction of product both for lateral phase separation in the gel state and for elimination of the lag phase is approximately 0.083. The simultaneous recordings of PLA2 activity and pyrene fluorescence show that the lateral rearrangement of lipids begins prior to and continues during the rapid activation process of PLA2. Two possible effects of lateral phase separation are that concentration of the protein in the product-rich regions promotes putative dimerization or that formation of phase interface regions promotes enzyme activation.

Enzyme Activation↗

PubMed2004

Surface phase separation in complex mixed adsorbing systems: an interface-bulk coupling effect.

The interfacial thermodynamics and structure of ternary mixtures of the type A+B+solvent are investigated. According to the Gibbs phase rule, the coupling between the bulk phase and the interfacial region-which is related to the reversibility of the adsorption of the corresponding species-is a determinant as to whether phase separation can be observed at the interface. For an n-component adsorbing solution, at least one of the species has to adsorb irreversibly over the experimental time scales in order not to fix more intensive variables than those required to observe surface phase separation. We present results for a lattice model planar interface consisting of the ternary mixture A+B+solvent. The solvent molecules and the type A molecules have fixed chemical potentials at the interface since they are equilibrated with a bulk solution. In contrast, the type B molecules are irreversibly adsorbed at the interface and do not equilibrate with the bulk. Mean-field theory is compared with Monte Carlo simulation. Interestingly, the spinodal line in the interaction-composition plane shows a reentrant on the B-rich phase side. We discuss the implications of these results for surface phase separation of adsorbing mixtures of proteins and low-molecular-weight surfactants.

Journal Article↗

PubMed1977

A unitary theory of anesthesia based on lateral phase separations in nerve membranes.

This paper relates research on anesthetic effects on lipid membrane systems to mechanisms of neural function. A unitary theory of anesthesia based on anesthetic-induced changes in fluid-solid-phase separations in the lipid region of nerve membranes is presented. It is suggested that anesthetics act by fluidizing nerve membranes to a point where critical lipid regions no longer contain phase separations. As a consequence, the membranes are less able to facilitate the conformational changes in proteins that may be the basis for such membrane events as ion gating, synaptic transmitter release, and transmitter binding to receptors. It is proposed that the anesthetic-modified phase separation behavior of the membrane may alter neural function by a combination of the following effects: inhibition of conformational changes of intrinsic membrane proteins; prevention of the association of protein subunits to form polymeric ion channels; depression of transmitter release by preventing fusion of vesicles containing synaptic transmitter with the membrane of the presynaptic terminal.

Anesthesia, Inhalation↗

PubMed1985

Phase separation in Triton X-114 of antigens of transmission blocking immunity in Plasmodium gallinaceum.

The distribution of proteins of mosquito midgut forms of Plasmodium gallinaceum in the detergent-free (aqueous) and detergent-enriched phases was studied using a phase separation technique in Triton X-114. Of the three surface proteins on gametes and newly fertilized zygotes (240, 56, and 54 kDa) immunoprecipitated by transmission blocking monoclonal antibodies, 240 kDa protein was recovered in the aqueous phase, whereas 56 and 54 kDa proteins were found preferentially in the detergent phase. The hydrophobic properties of the 56 and 54 kDa proteins were also shown by their strong tendency to interact with the lipid bilayers and a hydrophobic matrix phenyl-Sepharose. Monoclonal antibody IID3B3 immunoprecipitated all the three proteins from the whole Triton extract but in the phase-separated extracts reacted only with the 240 kDa protein in the aqueous phase and not with the 56 and 54 kDa doublet in the detergent phase. In Western blot analysis also monoclonal antibody IID3B3 reacted only with the 240 kDa protein. The 240 kDa protein in the aqueous phase was retained by monoclonal antibody IID3B3 linked to Sepharose 4B beads and could be eluted either with 0.1 M acetic acid or 50 mM diethylamine. The 56 and 54 kDa doublet in the detergent phase could be bound to and eluted from Sepharose 4B beads-linked monoclonal antibody IID4 or rabbit anti-male P. gallinaceum gamete serum. Two stage-specific glycoproteins of 26 and 28 kDa on the surface of ookinetes of P. gallinaceum were also separated in the detergent phase following Triton X-114 extraction. Phase separation in Triton X-114 offers a simple approach to the separation of a select group of proteins from the bulk of the cellular proteins.

Animals↗

PubMed1986

Demonstration of the amphiphilic character of hormone-sensitive lipase by temperature-induced phase separation in Triton X-114 and charge-shift electrophoresis.

Temperature-induced phase separation in Triton X-114 (Bordier, C. (1981) J. Biol. Chem. 256, 1604-1607) and charge-shift electrophoresis (Helenius, A., and Simons, K. (1977) Proc. Natl. Acad. Sci. U. S. A. 74, 529-532) were used to examine the amphiphilic character of hormone-sensitive lipase, purified from rat adipose tissue. In contrast to ATP-citrate lyase, a reference hydrophilic protein, the lipase was shown to partition predominantly (approximately 80%) into the detergent-rich phase upon phase separation in Triton X-114. Furthermore, its electrophoretic mobility was markedly shifted anodally and cathodally upon charge-shift electrophoresis in the presence of sodium taurodeoxycholate and cetyltrimethylammonium bromide, respectively. The results demonstrate that hormone-sensitive lipase possesses detergent-binding hydrophobic domain(s) and exhibits the same amphiphilicity as typical intrinsic membrane proteins.

ATP Citrate (pro-S)-Lyase↗

PubMed2005

Convection-driven pattern formation in phase-separating binary fluids.

Using a thermal-lattice Boltzmann model, we examine the rich phase behavior that develops when partially miscible fluids evolve in the presence of a vertical temperature gradient, which encompasses the critical temperature T(c) of the mixture. In particular, a binary AB fluid is confined between two plates in a gravitational field. The upper plate is fixed below T(c) and hence, the nearby fluid phase separates into A-rich and B-rich domains. The lower plate is fixed above the temperature T(c), and the surrounding fluid is in the homogeneous phase. A coupling between convection (driven by the temperature gradient) and phase separation gives rise to unique pattern formation. A number of regimes are identified: regularly spaced stripes, convective steady-state columns, the periodic disturbance of these columns, and finally, chaotic dripping from the upper surface. These results highlight dynamical behavior in partially miscible mixtures.

Journal Article↗

PubMed2000

Consequences of phase separation in cytoplasm.

Solutions of structurally different macromolecules, when mixed, above certain concentrations, tend to phase separate. The high concentration and diversity of proteins present in the liquid phase of cytoplasm, together with the phenomena which accompany macromolecular crowding, appear to meet the requirements for multiphase separation. The resulting cytoplasmic phase compartments, bounded by interfaces and/or other intracellular surfaces, would provide a dynamic, three-dimensional organizational structure. Based on the known physicochemical properties of aqueous phase systems and the partitioning behavior of biomaterials in them, such an organization could account for numerous phenomena observed in cytoplasm, such as its microcompartmentation. Aqueous phase separation is an attractive model for the liquid phase of cytoplasm because it comprises not only a structure (phase compartments, interfaces) for constraining biomaterials but also a mechanism (partitioning) for translocating them to other sites.

Animals↗

PubMed2006

Thick silica gel coatings on methylsilsesquioxane monoliths using anisotropic phase separation.

Silica gel coatings on methyltrimethoxysilane (MTMS)-derived monoliths have been studied using wetting transition. Wetting transition is observed in a small confined space, where a coating solution phase-separates into a well-coarsened dimension, making all the phase-separating polymerizing silica phase dynamically flow onto the existing surface of a mold. Bulk coating experiments have shown reductions of both macropore volume and diameter due to the coated layer. Comparing HPLC efficiencies of the coated monolith with those of the non-coated MTMS monolith revealed that the retention factors drastically increased in both normal- and reversed-phase modes. This is attributed to the existence of considerable amounts of accessible micropores left inside the coated layer, where analyte molecules travel and adsorb for a considerable period of time.

Journal Article↗

PubMed2001

Investigation of phase separation in Nd3+ doped ternary sodium borosilicate glasses by optical spectroscopy.

Two glasses doped with 1 mol% Nd2O3 and batch compositions inside the miscibility gap of the ternary Na2O-B2O3-SiO2 system were prepared by rapid quenching of the 1,400 degrees C melts. Phase separation was induced by heat-treatment at 600 degrees C for different exposure times and monitored qualitatively by an observation of Rayleigh scattering. The 4I(9/2-->P(1/2) transition of Nd3+ around 23300 cm(-1) recorded for the quenched samples without heat-treatment was used to demonstrate that submicroscopic phase separation in the doped glasses occurs instantaneously upon quenching. The effect of the Nd3+ concentration on this submicroscopic phase separation was investigated.

Biophysical Phenomena↗

PubMed2006

Phase separation in superoxygenated La2-xSrxCuO4+y.

The complex interplay between superconducting and magnetic phases remains poorly understood. Here, we report on the phase separation of doped holes into separate magnetic and superconducting regions in superoxygenated La(2-x)Sr(x)CuO(4+y), with various Sr contents. Irrespective of Sr-doping, excess oxygen raises the superconducting onset to 40 K with a coexisting magnetic spin-density wave that also orders near 40 K in each of our samples. The magnetic region is closely related to the anomalous, 1/8-hole-doped magnetic versions of La(2)CuO(4), whereas the superconducting region is optimally doped. The two phases are probably the only truly stable ground states in this region of the phase diagram. This simple two-component system is a candidate for electronic phase separation in cuprate superconductors, and a key to understanding seemingly conflicting experimental observations.

Journal Article↗

PubMed1999

Phase-separated composite films for liquid crystal displays

A method of preparing liquid crystal devices by phase separation of liquid crystal from its solution in a prepolymer, which results in adjacent layers of liquid crystal and polymer, is described. Liquid crystals in these phase-separated composite films exhibit electro-optical properties not observed in devices prepared by conventional methods, polymer dispersion, or polymer-stabilization methods. Devices incorporating ferroelectric liquid crystals have gray scale and switch 100 times faster at low fields than conventional surface-stabilized devices. This method makes it possible to prepare devices with liquid crystal film thickness comparable to optical wavelengths.

Journal Article↗

PubMed1991

Binary-liquid phase separation of lens protein solutions.

We have determined the coexistence curves (plots of phase-separation temperature T versus protein concentration C) for aqueous solutions of purified calf lens proteins. The proteins studied, calf gamma IIIa-, gamma IIIb-, and gamma IVa-crystallin, have very similar amino acid sequences and three-dimensional structures. Both ascending and descending limbs of the coexistence curves were measured. We find that the coexistence curves for each of these proteins and for gamma II-crystallin can be fit, near the critical point, to the function /(Cc-C)/Cc/ = A [(Tc - T)/Tc]beta, where beta = 0.325, Cc is the critical protein concentration in mg/ml, Tc is the critical temperature for phase separation in K, and A is a parameter that characterizes the width of the coexistence curve. We find that A and Cc are approximately the same for all four coexistence curves (A = 2.6 +/- 0.1, Cc = 289 +/- 20 mg/ml), but that Tc is not the same. For gamma II- and gamma IIIb-crystallin, Tc approximately 5 degrees C, whereas for gamma IIIa- and gamma IVa-crystallin, Tc approximately 38 degrees C. By comparing the published protein sequences for calf, rat, and human gamma-crystallins, we postulate that a few key amino acid residues account for the division of gamma-crystallins into low-Tc and high-Tc groups.

Animals↗