Neutron reflectivity experiments on the interface of pure D2O against thin films of perdeuterated polystyrene (d-PS) spin-coated onto silicon blocks were performed to study the intrinsic structure of the interface of water against hydrophobic substrates. The experiments reveal nonvanishing scattering contrast at the polymer/water interface, although the two materials (d-PS and D2O) have closely similar scattering length densities. Organic (nondeuterated) contaminants or macroscopic air bubbles trapped at the polymer/water interface can be ruled out as the origin of this observation. From a systematic study of this system, it is concluded that the source of the nonvanishing contrast is a depletion of water in the boundary layer against the hydrophobic surface. It is conjectured that this depletion layer represents a precursor layer of submicroscopic gas bubbles recently observed by Tyrrell and Attard. The existence of such gas nanobubbles in the present system is confirmed by atomic force microscopy (AFM) of the surface of d-PS coatings in contact with bulk water. The thickness of the precursor gas layer as determined by neutron reflectometry is 2−5 nm, depending on the level of air saturation of the water sample and on the time elapsed after contacting it with the hydrophobic surface.

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Dielectric spectroscopy is based on the response of the permanent dipoles to a driving electric field. The phospholipid membrane systems of dimyristoylphosphatidylcholine and dioleoylphosphatidylcholine can be prepared as samples of multilamellar liposomes with a well known amount of interlamellar water. For optimal resolution in dielectric spectroscopy one has to design the experimental set-up so that the direction of the permanent headgroup dipole moment is mostly parallel to the field vector of the external radio frequency (rf) electric field in this layered system. A newly developed coaxial probe technique makes it possible to sweep the measuring frequency between 1 and 1000 MHz in the temperature range 286-323 K. The response yields both the dispersion (epsilon') and the absorption part (epsilon") of the complex dielectric permittivity, which are attributed to the rotational diffusions of the zwitterionic phosphatidylcholine headgroup and the hydration water, respectively. Although the contributions of the headgroup and the hydration dipole moments to the dielectric relaxation are found to be situated close together, we succeeded in separating them. In the language of the Debye description, we propose to assign the lower frequency portion of the signal response to the relaxation contributed by the headgroups. The respective relaxation frequency is a discrete value in the range of 15-100 MHz and it shows normal temperature dependence. The contribution of the hydration water molecules exhibits a similar behavior in the range of 100-500 MHz but with the attributed relaxation frequency as the center of an asymmetric distribution of frequencies in analogy to simulation models known from the literature. Activation energies are derived for each of these relaxation processes from the Arrhenius plots of the temperature-dependent relaxation frequencies.

Cardiolipins (CLs) are important biologically for their unique role in biomembranes that couple phosphorylation and electron transport like bacterial plasma membranes, chromatophores, chloroplasts and mitochondria. CLs are often tightly coupled to proteins involved in oxidative phosphorylation. The first step in understanding the interaction of CL with proteins is to obtain the pure CL structure, and the structure of mixtures of CL with other lipids. In this work we use a variety of techniques to characterize the fluid phase structure, material properties and thermodynamics of mixtures of dimyristoylphosphatidylcholine (DMPC) with tetramyristoylcardiolipin (TMCL), both with 14-carbon chains, at several mole percentages. X-ray diffuse scattering was used to determine structure, including bilayer thickness and area/lipid, the bending modulus, KC, and Sxray, a measure of chain orientational order. Our results reveal that TMCL thickens DMPC bilayers at all mole percentages, with a total increase of ~6 Å in pure TMCL, and increases AL from 64 Å2 (DMPC at 35°C) to 109 Å2 (TMCL at 50°C). KC increases by ~50%, indicating that TMCL stiffens DMPC membranes. TMCL also orders DMPC chains by a factor of ~2 for pure TMCL. Coarse grain molecular dynamics simulations confirm the experimental thickening of 2 Å for 20 mol% TMCL and locate the TMCL headgroups near the glycerol-carbonyl region of DMPC; i.e., they are sequestered below the DMPC phosphocholine headgroup. Our results suggest that TMCL plays a role similar to cholesterol in that it thickens and stiffens DMPC membranes, orders chains, and is positioned under the umbrella of the PC headgroup. CL may be necessary for hydrophobic matching to inner mitochondrial membrane proteins. Differential scanning calorimetry, Sxray and CGMD simulations all suggest that TMCL does not form domains within the DMPC bilayers. We also determined the gel phase structure of TMCL, which surprisingly displays diffuse X-ray scattering, like a fluid phase lipid. AL = 40.8 Å2 for the ½TMCL gel phase, smaller than the DMPC gel phase with AL = 47.2 Å2, but similar to AL of DLPE = 41 Å2, consistent with untilted chains in gel phase TMCL.

Neutron reflectivity was applied to monitor in situ the adsorption of small unilamellar phospholipid vesicles on a solid bare hydrophilic Si interface. The obtained reflectivity curves are consistent with the rupture and fusion model for the adsorption of phosphatidylcholine vesicles to solid interfaces. The results show details of the adsorbed bilayer system at ångström resolution and indicate the presence of a thin thick water leaflet that separates the bilayer from the Si surface. The resolved structural details provide the basis for further investigation of processes such as adsorption and penetration of peptides and proteins towards the supported bilayer at high resolution.