The structure of the Staphylococcus aureus α-hemolysin pore has been determined to 1.9 A resolution. Contained within the mushroom-shaped homo-oligomeric heptamer is a solvent-filled channel, 100 A in length, that runs along the sevenfold axis and ranges from 14 A to 46 A in diameter. The lytic, transmembrane domain comprises the lower half of a 14-strand antiparallel β barrel, to which each protomer contributes two β strands, each 65 A long. The interior of the β barrel is primarily hydrophilic, and the exterior has a hydrophobic belt 28 A wide. The structure proves the heptameric subunit stoichiometry of the α-hemolysin oligomer, shows that a glycine-rich and solvent-exposed region of a water-soluble protein can self-assemble to form a transmembrane pore of defined structure, and provides insight into the principles of membrane interaction and transport activity of β barrel pore-forming toxins.

https://science.sciencemag.org/content/sci/274/5294/1859.full.pdf

Structure of Staphylococcal α-Hemolysin, a Heptameric Transmembrane Pore

Molecular dynamics simulations of a fluid bilayer of dipalmitoylphosphatidylcholine at full hydration, constant pressure, and constant temperature.

Cholesterol and the cell membrane.

Proton and carbon-13 nmr spectra of unsonicated lipid bilayers and biological membranes are generally dominated by strong proton–proton and proton–carbon dipolar interactions. As a result the spectra contain a large number of overlapping resonances and are rather difficult to analyse. Nevertheless, important information on the structure and dynamic behaviour of lipid systems has been provided by these techniques (Wennerstrom & Lindblom, 1977).

/pdf/deuterium-magnetic-resonance-theory-and-application-to-lipid-3a016gcplt.pdf

Deuterium magnetic resonance: theory and application to lipid membranes.

Deuterium nuclear magnetic resonance spectroscopy and differential scanning calorimetry are used to map the phase boundaries of mixtures of cholesterol and chain-perdeuteriated 1,2-dipalmitoyl-sn- glycero-3-phosphocholine at concentrations from 0 to 25 mol % cholesterol. Three distinct phases can be identified: the La or liquid-crystalline phase, the gel phase, and a high cholesterol concentration phase, which we call the 0 phase. The liquid-crystalline phase is characterized by highly flexible phospholipid chains with rapid axially symmetric reorientation; the gel phase has much more rigid lipid chains, and the motions are no longer axially symmetric on the 2H NMR time scale; the 0 phase is characterized by highly ordered (rigid) chains and rapid axially symmetric reorientation. In addition, we identify three regions of two-phase coexistence. The first of these is a narrow La/gel-phase coexistence region lying between 0 and about 6 mol % cholesterol at temperatures just below the chain-melting transition of the pure phospho- lipid/water dispersions, at 37.75 OC. The dramatic changes in the *H NMR line shape which occur on passing through the phase transition are used to map out the boundaries of this narrow two-phase region. The boundaries of the second two-phase region are determined by 2H NMR difference spectroscopy, one boundary lying near 7.5 mol 5% cholesterol and running from 37 down to at least 30 OC; the other boundary lies near 22 mol 5% cholesterol and covers the same temperature range. Within this region, the gel and /3 phases coexist. As the temperature is lowered below about 30 "C, the phospholipid motions reach the intermediate time scale regime of 2H NMR so that spectral subtractions become difficult and unreliable. The third two-phase region lies above 37 OC, beginning at a eutectic point somewhere between 7.5 and 10 mol % cholesterol and ending at about 20 mol %. In this region, the La and /3 phases are in equilibrium. The boundaries for this region are inferred from differential scanning calorimetry traces, for the boundary between the La- and the two-phase region, and from a dramatic sharpening of the NMR peaks on crossing the boundary between the two-phase region and the &phase region. In this region, the technique of difference spectroscopy fails, presumably because the diffusion rate in both the La- and P-phase domains is so rapid that phospholipid molecules exchange rapidly between domains on the experimental time scale.

D.L. Worcester

Papers

Structural analysis of hydrated egg lecithin and cholesterol bilayers I. X-ray diffraction