Mu-Ping Nieh

Fluid phase lipid areas and bilayer thicknesses of commonly used phosphatidylcholines as a function of temperature.

The structural parameters of fluid phase bilayers composed of phosphatidylcholines with fully saturated, mixed, and branched fatty acid chains, at several temperatures, have been determined by simultaneously analyzing small-angle neutron and X-ray scattering data. Bilayer parameters, such as area per lipid and overall bilayer thickness have been obtained in conjunction with intrabilayer structural parameters (e.g. hydrocarbon region thickness). The results have allowed us to assess the effect of temperature and hydrocarbon chain composition on bilayer structure. For example, we found that for all lipids there is, not surprisingly, an increase in fatty acid chain trans-gauche isomerization with increasing temperature. Moreover, this increase in trans-gauche isomerization scales with fatty acid chain length in mixed chain lipids. However, in the case of lipids with saturated fatty acid chains, trans-gauche isomerization is increasingly tempered by attractive chain-chain van der Waals interactions with increasing chain length. Finally, our results confirm a strong dependence of lipid chain dynamics as a function of double bond position along fatty acid chains.

Applications of neutron and X-ray scattering to the study of biologically relevant model membranes

Scattering techniques, in particular electron, neutron and X-ray scattering have played a major role in elucidating the static and dynamic structure of biologically relevant membranes. Importantly, neutron and X-ray scattering have evolved to address new sample preparations that better mimic biological membranes. In this review, we will report on some of the latest model membrane results, and the neutron and X-ray techniques that were used to obtain them.

Morphology of fast-tumbling bicelles: a small angle neutron scattering and NMR study

Bilayered micelles, or bicelles, which consist of a mixture of long- and short-chain phospholipids, are a popular model membrane system. Depending on composition, concentration, and temperature, bicelle mixtures may adopt an isotropic phase or form an aligned phase in magnetic fields. Well-resolved (1)H NMR spectra are observed in the isotropic or so-called fast-tumbling bicelle phase, over the range of temperatures investigated (10-40 degrees C), for molar ratios of long-chain lipid to short-chain lipid between 0.20 and 1.0. Small angle neutron scattering data of this phase are consistent with the model in which bicelles were proposed to be disk-shaped. The experimentally determined dimensions are roughly consistent with the predictions of R.R. Vold and R.S. Prosser (J. Magn. Reson. B 113 (1996)). Differential paramagnetic shifts of head group resonances of dimyristoylphosphatidylcholine (DMPC) and dihexanoylphosphatidylcholine (DHPC), induced by the addition of Eu(3+), are also consistent with the bicelle model in which DHPC is believed to be primarily sequestered to bicelle rims. Selective irradiation of the DHPC aliphatic methyl resonances results in no detectable magnetization transfer to the corresponding DMPC methyl resonances (and vice versa) in bicelles, which also suggests that DHPC and DMPC are largely sequestered in the bicelle. Finally, (1)H spectra of the antibacterial peptide indolicidin (ILPWKWPWWPWRR-NH(2)) are compared, in a DPC micellar phase and the above fast-tumbling bicellar phases for a variety of compositions. The spectra exhibit adequate resolution and improved dispersion of amide and aromatic resonances in certain bicelle mixtures.

SANS Study on the Effect of Lanthanide Ions and Charged Lipids on the Morphology of Phospholipid Mixtures

The structural phase behavior of phospholipid mixtures consisting of short-chain (dihexanoyl phosphatidylcholine) and long-chain lipids (dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylglycerol), with and without lanthanide ions was investigated by small-angle neutron scattering (SANS). SANS profiles were obtained from 10 degrees C to 55 degrees C using lipid concentrations ranging from 0.0025 g/ml to 0.25 g/ml. The results reveal a wealth of distinct morphologies, including lamellae, multi-lamellar vesicles, unilamellar vesicles, and bicellar disks.

“Bicellar” Lipid Mixtures as used in Biochemical and Biophysical Studies

Over the past decade “bicellar” lipid mixtures composed of the long-chain dimyristoyl phosphatidylcholine (DMPC) and the short-chain dihexanoyl PC (DHPC) molecules have emerged as a powerful medium for studying membrane associated, biologically relevant macromolecules and assemblies. Depending on temperature, lipid concentration and composition these lipid mixtures can assume a variety of morphologies, some of them alignable in the presence of a magnetic field. This article will examine the biophysical studies that have elucidated the various morphologies assumed by these lipid mixtures, and their use in the biochemical studies of biomolecules.

Chain Conformation of a New Class of PEG-Based Thermoresponsive Polymer Brushes Grafted on Silicon as Determined by Neutron Reflectometry

The thermoresponsive PEG-based copolymer poly[2-(2-methoxyethoxy)ethyl methacrylate-co-oligo(ethylene glycol) methacrylate) (P(MEO(2)MA-co-OEGMA)] was grafted onto a silicon wafer, and its chain conformation in aqueous solution was studied by neutron reflectometry. The effects of temperature and salt concentration on the polymers conformation were evaluated. With increasing temperature, it was found that the polymer brushes underwent a transition from an extended state to a compressed state, and eventually a collapsed state above the lower critical solution temperature. The presence of salt significantly affected the well-extended brushes but had little effect on compressed and collapsed brushes. This PEG-based thermoresponsive surface exhibited good protein adsorption resistance. Interestingly, extended and collapsed brushes showed the same level of protein repulsion, something that was not expected.

Effect of cations on the structure of bilayers formed by lipopolysaccharides isolated from Pseudomonas aeruginosa PAO1.

The asymmetric outer membrane of Gram-negative bacteria contains lipopolysaccharides (LPSs) which contribute significantly to the bacteriums surface properties and play a crucial role in regulating membrane permeability. We report on neutron diffraction studies performed on aligned, self-assembled bilayers of Na-, Ca-, and Mg-salt forms of LPS isolated from Pseudomonas aeruginosa PAO1. From the one-dimensional neutron scattering length density profiles we find that water penetrates Ca2+-LPS bilayers to a lesser extent than either Na+- or Mg2+-LPS bilayers. This differential water penetration could have implications as to how small molecules permeate the outer membrane of Gram-negative bacteria and, possibly, how nonlamellar phases are formed.

Spontaneously Formed Monodisperse Biomimetic Unilamellar Vesicles: The Effect of Charge, Dilution, and Time

Using small-angle neutron scattering and dynamic light scattering, we have constructed partial structural phase diagrams of lipid mixtures composed of the phosphatidylcholines dimyristoyl and dihexanoyl doped with calcium ions (Ca2+) and/or the negatively charged lipid, dimyristoyl phosphatidylglycerol (DMPG). For dilute solutions (lipid concentration < or =1 wt %), spontaneously forming unilamellar vesicles (ULVs) were found, and their polydispersity was determined to be approximately 20%. The stability of the Ca2+- or DMPG-doped ULVs was monitored over a period of 4 days and their structural parameters (e.g., average outer radius, ) were found to be insensitive to the lipid concentration (Clp). However, doping the dimyristoyl/dihexanoyl system with both Ca2+ and DMPG resulted in ULVs whose was found to be Clp dependent. The of DMPG-doped ULVs remained unchanged over an extended period of time (at least 4 days), a good indication of their stability.

The Functional Significance of Lipid Diversity: Orientation of Cholesterol in Bilayers Is Determined by Lipid Species

The chemical diversity of lipids and their complex arrangements in supramolecular assemblies are in stark contrast to our previous notions of them as passive structural components. For example, in plasma membranes, sphingolipids are primarily located in the outer monolayer, whereas unsaturated phospholipids are more abundant in the inner leaflet. Our recent results offer a direct contribution to the importance of lipid diversity in biological membranes. We have studied the location of cholesterol within polyunsaturated fatty acid (PUFA) bilayers doped with different amounts of monounsaturated (POPC) or disaturated (DMPC) lipids. Using deuterium labeling and neutron diffraction, we have found that in PUFA bilayers, cholesterol can be flipped from its known position in the bilayer center to its commonly assumed upright orientation simply by varying the amount of POPC. Although it takes approximately 50 mol % POPC to flip cholesterol in PUFA bilayers, the same effect is achieved with only 5 mol % DMPC, elegantly emphasizing cholesterols affinity for saturated chains. It also suggests that the presence of PUFA in the inner leaflet of a cellular bilayer may enhance the transfer of cholesterol to the outer layer, potentially modifying raft composition and the local function of a membrane.

A fluorescent polymer film with self-assembled three-dimensionally ordered nanopores: preparation, characterization and its application for explosives detection

The sensitive and rapid detection of nitroaromatic explosives typically requires sophisticated sensor materials. We demonstrate here how a simple dip-coating process of a mixture of polystyrene and the fluorophore pyrene onto a glass slide generates a self-assembled fluorescent nanostructured film expressing regular breath-figure nanopores. Morphology investigation reveals that the fluorescent polymer films consist of a high-density, three-dimensional nanoporous array of holes, allowing the fluorescence of this material to be rapidly and selectively quenched by nitroaromatic vapors. The morphology of the polymer was controlled by variation of the dip-coating parameters and the ratio of polystyrene to pyrene. This ratio also controls the fluorescence quenching efficiency of the material. We demonstrate the possible molecular origins of this through structural XRD studies as well as investigations of the electronic structure (optical properties, band gap and conduction band determinations) of the polymer film. Our results identify a novel high performance form of an otherwise known explosive-sensing material. Most importantly, the findings point toward a general method for the facile realization of well-defined three-dimensional high surface sensor materials with optimized electronic properties.