Pressure perturbation calorimetry of unfolded proteins.

We report the application of pressure perturbation calorimetry (PPC) to study unfolded proteins. Using PPC we have measured the temperature dependence of the thermal expansion coefficient, α(T), in the unfolded state of apocytochrome C and reduced BPTI. We have shown that α(T) is a nonlinear function and decreases with increasing temperature.

Monitoring detergent-mediated solubilization and reconstitution of lipid membranes by isothermal titration calorimetry.

The solubilization and reconstitution of biological or liposomal membranes by detergents and biomolecules with detergent-like properties play a major role for technical applications (e.g., the isolation of membrane proteins) and biological phenomena (of, e.

Modeling the micellization behavior of mixed and pure n-alkyl-maltosides.

The micellization behavior of a series of n-alkyl-maltosides in aqueous solution was studied by isothermal titration calorimetry (ITC) and dynamic light scattering (DLS) at 25 degrees C. Demicellization experiments were conducted with single component micelles of octyl (OM), nonyl (NM), decyl (DM), undecyl (UM), and dodecyl (lauryl, LM) maltoside and binary mixtures of LM with OM, NM, DM and UM, respectively. A model was derived on the basis of the pseudophase approximation to fit the complete demicellization curves.

Helix-coil transition of DNA monitored by pressure perturbation calorimetry.

We report the first use of pressure perturbation calorimetry (PPC) to characterize the heat-induced helix-coil transition of DNA polymers. The alternating copolymer poly[d(A-T)] was studied in aqueous solutions containing 5.2 and 18.

Additive action of two or more solutes on lipid membranes.

A wide variety of biological processes, pharmaceutical applications, and technical procedures is based on the combined action of two or more soluble compounds to perturb, permeabilize, or lyse biological membranes. Here we present a general model describing the additive action of solutes on the properties of membranes or micelles. The onset and completion of membrane solubilization induced by two surfactants (lauryl maltoside, with nonyl maltoside, octyl glucoside, or CHAPS, respectively) are very well described by our model on the basis of their individual partition coefficients, cmc's, and critical mole ratios R e sat and R e sol as detected by isothermal titration calorimetry.

Uptake and release protocol for assessing membrane binding and permeation by way of isothermal titration calorimetry.

The activity of many biomolecules and drugs crucially depends on whether they bind to biological membranes and whether they translocate to the opposite lipid leaflet and trans aqueous compartment. A general strategy to measure membrane binding and permeation is the uptake and release assay, which compares two apparent equilibrium situations established either by the addition or by the extraction of the solute of interest. Only solutes that permeate the membrane sufficiently fast do not show any dependence on the history of sample preparation.

Nonideal mixing in multicomponent lipid/detergent systems.

A detailed understanding of the mixing properties of membranes to which detergents are added is mandatory for improving the application and interpretation of detergent based protein or lipid extraction assays. For Triton X-100 (TX-100), a nonionic detergent frequently used in the process of solubilizing and purifying membrane proteins and lipids, we present here a detailed study of the mixing properties of binary and ternary lipid mixtures by means of high-sensitivity isothermal titration calorimetry (ITC). To this end the partitioning thermodynamics of TX-100 molecules from the aqueous phase to lipid bilayers composed of various mixtures of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), egg-sphingomyelin (SM), and cholesterol (cho) are characterized.

Gradual change or phase transition: characterizing fluid lipid-cholesterol membranes on the basis of thermal volume changes.

Cholesterol has been reported to govern biomembrane permeability, elasticity, and the formation of lipid rafts. There has been a controversy whether binary lipid-cholesterol membranes should better be described in terms of a phase separation (liquid-ordered and liquid-disordered phases) or of gradual changes in largely homogeneous membranes. We present a new approach for detecting and characterizing phase equilibria in colloidal dispersions using pressure perturbation calorimetry (PPC).

Thermodynamic comparison of the interactions of cholesterol with unsaturated phospholipid and sphingomyelins.

A comparative analysis of the interaction of cholesterol (Chol) with palmitoyl-oleoyl-phosphatidylcholine (POPC) and sphingomyelins (SM) was performed in largely homogeneous, fluid-phase membranes at 50 degrees C. To this end, three independent assays for isothermal titration calorimetry were applied to POPC/SM/Chol mixtures. Cholesterol is solubilized by randomly methylated-beta-cyclodextrin and the uptake of Chol into (or release from) large unilamellar vesicles is measured.

A quantitative model describing the selective solubilization of membrane domains.

The classical three-stage model of membrane solubilization, including mixed membranes, membrane-micelle coexistence, and mixed micelles, is not applicable to demixed, domain-forming membranes and must, therefore, fail to describe the phenomenon of detergent-resistant membranes (DRMs). In lack of a quantitative model, it has often been assumed that ordered, detergent-depleted domains are inert, whereas fluid domains are solubilized. We establish a quantitative model based on equilibrium thermodynamics that is analogous to the three-stage model but comprises three components (two lipids and one detergent) in four phases (liquid-ordered and liquid-disordered membranes, micelles, and detergent in aqueous solution).

Interactions of cholesterol with lipid membranes and cyclodextrin characterized by calorimetry.

Interactions of cholesterol (cho) with different lipids are commonly believed to play a key role in the formation of functional domains in membranes. We introduce a novel approach to characterize cho-lipid interactions by isothermal titration calorimetry. Cho is solubilized in the aqueous phase by reversible complexation with methyl-beta-cyclodextrin (cyd).

Structural, volumetric, and thermodynamic characterization of a micellar sphere-to-rod transition.

The thermotropic sphere-to-rod transition of nonionic surfactants was characterized in terms of a large set of parameters: the transition temperature and width, the partial volume, coefficient of thermal volume expansion, enthalpy, isobaric heat capacity, and structural parameters, such as radius of gyration and hydrodynamic radius. Data were recorded as a function of concentration of surfactants in H2O and in D2O. To this end, pressure perturbation calorimetry (PPC), small angle neutron scattering (SANS), dynamic light scattering (DLS), differential scanning calorimetry (DSC), and isothermal titration calorimetry (ITC) were applied in a study of aqueous solutions containing myristyl, tridecyl, and lauryl maltoside and heptaethyleneglycoltetradecyl ether (C14EO7).