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Probing self assembly in biological mixed colloids by SANS, deuteration, and molecular manipulation.

Small-angle neutron scattering was used to obtain information on the form and molecular arrangement of particles in mixed colloids of bile salts with phosphatidylcholine, and bile salts with monoolein. Both types of systems showed the same general characteristics. The particle form was highly dependent on total lipid concentration. At the highest concentrations the particles were globular mixed micelles with an overall size of 50A. As the concentration was reduced the mixed micelles elongated, becoming rodlike with diameter about 50A. The rods had a radial core-shell structure in which the phosphatidylcholine or monoolein fatty tails were arranged radially to form the core with the headgroups pointing outward to form the shell. The bile salts were at the interface between the shell and core with the hydrophilic parts facing outward as part of the shell. The lengths of the rods increased and became more polydispersed with dilution. At sufficiently low concentrations the mixed micelles transformed into single bilayer vesicles. These results give insight on the physiological function of bile and on the rules governing the self assembly of bile particles in the hepatic duct and the small intestine.

Bile Acids and Salts↗

Small angle scattering of cell nuclei.

Neutron and X-ray small angle scattering techniques have been applied to study chromatin structure inside different types of cell nuclei. Scattering from genetically inactive chicken erythrocyte nuclei exhibits a maximum at Q = 0.1-0.15 nm-1 which cannot be observed by studying isolated chromatin derived from the same kind of cells. In highly active transcribing rat liver nuclei such a nuclear pattern is absent. The radius of gyration of isolated "superbeads" was determined. It is discussed whether the characteristic maximum of the nuclei originates from this superstructural organisation of chromatin. Rat liver nuclei were fractionated on sucrose gradients in order to determine whether the absence of the extra maximum in scattering profiles of these nuclei is due to overlapping effects of different chromatin organisation in the various cell types of the liver. As compared to unfractionated nuclei no strong deviations in the scattering profiles of the fractions could be observed. Erythrocyte nuclei were dialysed in buffers differing in the ionic strength of monovalent cations. The typical maximum from the nuclei is shifted from 60 nm (very low salt concentration) to about 35 nm (physiological ionic strength) and is linearly proportional to the decreasing radius of the nuclei. In conclusion, chromatin structure inside the nucleus has a scattering maximum due to an ordered packing of the fibres which is absent in nuclei with high genetic activity.

Animals↗

Preparation and characterization of Boron carbide nanoparticles for use as a novel agent in T cell-guided boron neutron capture therapy.

Boron carbide nanoparticles are proposed as a system for T cell-guided boron neutron capture therapy. Nanoparticles were produced by ball milling in various atmospheres of commercially available boron carbide. The physical and chemical properties of the particles were investigated using transmission electron microscopy, photon correlation spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction, vibrational spectroscopy, gel electrophoresis and chemical assays and reveal profound changes in surface chemistry and structural characteristics. In vitro thermal neutron irradiation of B16 melanoma cells incubated with sub-100 nm nanoparticles (381.5 microg/g (10)B) induces complete cell death. The nanoparticles alone induce no toxicity.

Animals↗

The location of bound lipid in the lipovitellin complex.

The location of the bound lipid in the soluble lipoprotein lipovitellin has been determined by neutron crystallographic techniques. With the use of the contrast variation method, whereby the crystals are soaked in different H2O-D2O mixtures, the lipid has been found to occupy a large cavity in the protein whose structure had previously been determined by x-ray crystallography. The lipid appears to be bound in the form of a bilayer with the major protein-lipid interactions being hydrophobic and with the lipid headgroups projecting into the bulk solvent and into a solvent-filled space in the cavity.

Binding Sites↗

Finite-size effects do not reduce the repeat spacing of phospholipid multibilayer stacks on a rigid substrate.

Finite-size effects in stacks of phospholipid bilayers, in the fluid L alpha phase, are investigated using samples oriented on silicon substrates. Recently in this journal, such effects have been suggested as the probable cause of reduced lamellar repeat spacings in very thin samples made up of a few (<10) bilayers. Our systematic studies on samples of different thicknesses do not support this conclusion. At full hydration all samples are found to have the same repeat spacing, irrespective of their thickness. At lower hydrations, on the other hand, very thin samples, consisting of only a few bilayers, have a slightly larger spacing.

Biophysics↗

Halophilic malate dehydrogenase--a case history of biophysical investigations: ultracentrifugation, light-, X-ray- and neutron scattering.

Halophilic malate dehydrogenase (hMDH) from Haloarcula marismortui has been isolated, purified and characterized by biochemical and biophysical solution studies. A stabilization mechanism at extremely high concentrations of salt, based on the formation of co-operative hydrate bonds between the protein and hydrated salt ions, was suggested from thermodynamic analysis of native enzyme solutions. Recently the gene coding for hMDH was isolated and sequenced and an active enzyme cloned (F. Cendrin, J. Chroboczek, G. Zaccai, H. Eisenberg and M. Mevarech, unpublished work). A study of the crystal structure of hMDH in a high-salt physiological medium is in progress (O. Butbul-Dym & J. Sussman, personal communication). Here we discuss in depth implications of these recent developments on our earlier results.

Chemical Phenomena↗

Atomic string holography.

A new diffraction-channeling effect has been discovered, in which Kikuchi or channeling line patterns formed by high energy electrons, neutrons, and positrons are shown to break up into a series of annular disks if the crystal thickness traversed by the beam is small. The disks may be interpreted as Gabor in-line holograms of strings of atoms projected along the beam path. For electrons or positrons the patterns may be detected with little background by detecting characteristic x-ray emission from a thin film as a function of the diffraction conditions of a collimated, ionizing, high energy beam. Uses of the effect for structure determination and atomic-resolution lensless imaging are suggested.

Journal Article↗

Phase recovery and lensless imaging by iterative methods in optical, X-ray and electron diffraction.

Thomas Young's quantitative analysis of interference effects provided the confidence needed to revive the wave theory of light, and firmly established the concept of phase in optics. Phase plays a similarly fundamental role in matter-wave interferometry, for which the field-emission electron microscope provides ideal instrumentation. The wave-particle duality is vividly demonstrated by experimental 'Young's fringes' using coherent electron beams under conditions in which the flight time is less than the time between particle emission. A brief historical review is given of electron interferometry and holography, including the Aharonov-Bohm effect and the electron Sagnac interferometer. The simultaneous development of phase-contrast imaging at subnanometre spatial resolution has greatly deepened our understanding of atomic processes in biology, materials science and condensed-matter physics, while electron holography has become a routine tool for the mapping of electrostatic and magnetic fields in materials on a nanometre scale. The encoding of phase information in scattered farfield intensities is discussed, and non-interferometric, non-crystallographic methods for phase retrieval are reviewed in relationship to electron holography. Examples of phase measurement and diffraction-limited imaging using the hybrid input-output iterative algorithm are given, including simulations for soft X-ray imaging, and new experimental results for coherent electron and visible-light scattering. Image reconstruction is demonstrated from experimental electron and visible-light Fraunhofer diffraction patterns. The prospects this provides for lensless imaging using particles for which no lenses exist (such as neutrons, condensates, coherent atom beams and X-rays) are discussed. These new interactions can be expected to provide new information, perhaps, for example, in biology, with the advantage of less damage to samples.

Algorithms↗

Submicrometer coherent neutron beam production using a thin-film waveguide.

We have experimentally demonstrated that planar neutron waveguides can be used as resonant beam couplers to efficiently produce a coherent neutron line source with cross sections in the submicrometer range. The Fraunhofer far-field diffraction pattern of the first three resonance modes was measured and found to be in excellent agreement with the theoretical model. Our measurements confirm that an excited exiting mode is fully coherent in the direction perpendicular to the surface of the thin-film coupler and may therefore be used for applications of interest to a broad user community in biochemical and semiconductor nanosciences such as static and time-resolved coherent speckle experiments or phase-contrast imaging.

Journal Article↗

High-temperature phases of NaNbO3 and NaTaO3.

The high-temperature phases of the perovskites sodium niobate, NaNbO(3), and sodium tantalate, NaTaO(3), have been re-examined using the high-resolution powder diffractometer HRPD at the ISIS neutron spallation source; the two materials show the same sequence of phases with tilted octahedra. Diffraction patterns were measured every 5 K allowing structural changes with temperature within a single phase to be determined for the first time. Previous structure determinations within one phase had been performed at a single temperature only. The octahedra are tilted about pseudocubic <100> directions and are also deformed; the magnitude of the deformation is shown to be proportional to the square of the angle of tilt as expected from a phenomenological theory applied to such transitions. The structures of NaNbO(3) between 753 and 793 K and of NaTaO(3) below 758 K are not as reported in the literature.

Journal Article↗

The new HMI beamline MAGS: an instrument for hard X-ray diffraction at BESSY.

The Hahn-Meitner-Institute Berlin is operating the new hard X-ray diffraction beamline MAGS at the Berlin synchrotron radiation source BESSY. The beamline is intended to complement the existing neutron instrumentation at the Berlin Neutron Scattering Centre. The new beamline uses a 7 T multipole wiggler to produce photon fluxes in the 10(11)-10(12) photons s(-1) (100 mA)(-1) (0.1% bandwidth)(-1) range at energies from 4 to 30 keV at the experiment. It has active bendable optics to provide flexible horizontal and vertical focusing and to compensate the large heat load from the wiggler source. The experimental end-station consists of a six-circle Huber diffractometer which can be used with an additional (polarization) analyser and different sample environments. The beamline is intended for single-crystal diffraction and resonant magnetic scattering experiments for the study of ordering phenomena, phase transitions and materials science.

Equipment Design↗

Synchrotron and neutron techniques in biological crystallography.

Synchrotron radiation (SR) techniques are continuously pushing the frontiers of wavelength range usage, smaller crystal sample size, larger protein molecular weight and complexity, as well as better diffraction resolution. The new research specialism of probing functional states directly in crystals, via time-resolved Laue and freeze trapping structural studies, has been developed, with a range of examples, based on research stretching over some 20 years. Overall, SR X-ray biological crystallography is complemented by neutron protein crystallographic studies aimed at cases where much more complete hydrogen details are needed involving synergistic developments between SR and neutron Laue methods. A big new potential exists in harnessing genome databases for targeting of new proteins for structural study. Structural examples in this tutorial review illustrate new chemistry learnt from biological macromolecules.

Crystallography, X-Ray↗

Structural studies of plasma lipoproteins.

These studies with neutron small-angle scattering were designed to probe the location of the apoprotein of HDL on recombinant particles formed with phospholipids. The use of isotopic substitution with deuterium in the headgroup of the phospholipid allows the positions of the molecular components to be probed in more detail. These initial experiments point to the direction future structural studies must take. Reconstitution techniques together with compositional modification of native lipoproteins must be coupled with a combination of high accuracy x-ray and neutron scattering studies, the latter employing isotopic substitution with deuterium to highlight specific areas of the structure. This approach will undoubtedly provide a higher resolution picture of the molecular organization of plasma lipoproteins.

Apolipoproteins↗

Self-assembly of brome mosaic virus capsids. Kinetic study using neutron and X-ray solution scattering.

The self-assembly of brome mosaic virus capsid has been studied kinetically by means of X-ray and neutron scattering. It appears to be a very fast process: for the concentrations used (5 to 8 mg/ml) the forward scattering reaches 50% of its maximal value in less than one second. Further, the assembly seems to proceed through intermediate states whose nature is still speculative.

Capsid↗