Small-Angle Neutron Scattering at the Pulsed Reactor IBR-2: Current Status and Prospects

Crystallography Reports - Neutron diffraction studies on the small-angle neutron spectrometer YuMO (Joint Institute for Nuclear Research, Dubna), based on the IBR-2 pulsed reactor, have been...     

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 bacterium's 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.     

Curvature effect on the structure of phospholipid bilayers

High-resolution small-angle X-ray scattering (SAXS), complemented by small-angle neutron scattering (SANS) and dynamic light scattering (DLS) experiments, was used to study the effect of curvature on the bilayer structure of dioleoyl-phosphatidylcholine (DOPC) and dioleoyl-phosphatidylserine (DOPS) unilamellar vesicles (ULVs). Bilayer curvature, as a result of finite vesicle size, was varied as a function of vesicle radius and determined by DLS and SANS measurements. Unilamellarity of large DOPC ULVs was achieved by the addition of small amounts (up to 4 mol %) of the charged lipid, DOPS. A comparison of SANS data over the range of 0.02 < q <0.2 A-1 indicated no change in the overall bilayer thickness as a function of ULV diameter (620 to 1840 A). SANS data were corroborated by high-resolution (0.06 < q <0.6 A-1) SAXS data for the same diameter ULVs and data obtained from planar samples of aligned bilayers. Both the inner and outer leaflets of the bilayer were found to be indistinguishable. This observation agrees well with simple geometric models describing the effect of vesicle curvature. However, 1220-A-diameter pure DOPS ULVs form asymmetric bilayers whose structure can most likely be rationalized in terms of geometrical constraints coupled with electrostatic interactions, rather than curvature alone.     

Neutron diffraction study of Pseudomonas aeruginosa lipopolysaccharide bilayers

Lipopolysaccharides (LPSs) are a major class of macromolecules populating the surface of Gram-negative bacteria. They contribute significantly to the bacterium's surface properties and play a crucial role in regulating the permeability of its outer membrane. Here, we report on neutron diffraction studies performed on aligned, self-assembled bilayers of LPS isolated from Pseudomonas aeruginosa PAO1. This LPS system is comprised of a mixture of rough and smooth A-band and B-band LPS, similar to that naturally found in P. aeruginosa. Temperature scans were conducted at various levels of hydration, and the phases adopted by LPS, along with their corresponding transition temperatures, have been identified. Because of LPS's chemical heterogeneity, the gel-to-liquid-crystalline transition was continuous and not abrupt as commonly observed in single-component phospholipid systems. From the construction of one-dimensional scattering length density profiles, we find that water penetrates into the hydrocarbon region up to and including the center of liquid-crystalline LPS bilayers. This permeability to water also extends to bilayers in the continuous phase transition region and could have far-reaching implications as to how small molecules penetrate the outer membrane of Gram-negative bacteria.     

Erratum to: Small-Angle Neutron Scattering at the Pulsed Reactor IBR-2: Current Status and Prospects

Crystallography Reports - An Erratum to this paper has been published: https://doi.org/10.1134/S1063774522070513