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A A Chernov

Publications and source records attributed to A A Chernov.

13 recordsLinked to original sources

Diffraction and imaging study of imperfections of crystallized lysozyme with coherent X-rays.

Phase-contrast X-ray diffraction imaging and high-angular-resolution diffraction combined with phase-contrast radiographic imaging were employed to characterize defects and perfection of a uniformly grown tetragonal lysozyme crystal in the symmetric Laue case. The full-width at half-maximum (FWHM) of a 4 4 0 rocking curve measured from the original crystal was approximately 16.7 arcsec and imperfections including line defects, inclusions and other microdefects were observed in the diffraction images of the crystal. The observed line defects carry distinct dislocation features running approximately along the <1 1 0> growth front and have been found to originate mostly in a central growth area and occasionally in outer growth regions. Inclusions of impurities or formations of foreign particles in the central growth region are resolved in the images with high sensitivity to defects. Slow dehydration led to the broadening of a fairly symmetric 4 4 0 rocking curve by a factor of approximately 2.6, which was primarily attributed to the dehydration-induced microscopic effects that are clearly shown in X-ray diffraction images. The details of the observed defects and the significant change in the revealed microstructures with drying provide insight into the nature of imperfections, nucleation and growth, and the properties of protein crystals.

Animals↗

[The effect of heterozygosity for marker loci on recombination fraction in adjacent chromosomal regions in maize].

Analysis of maize F2 plants revealed a decrease in the recombination fraction (rf) between the marker loci in case of heterozygosity of adjacent chromosomal regions. The relative decrease in rf was positively associated with the length of the heterozygous region. Heterozygosity had no effect on rf in other chromosomes. These results can be explained by defective pairing of heterozygous chromosomal regions in meiosis.

Chromosomes, Plant↗

Surface relaxation in protein crystals.

Surface x-ray diffraction measurements were performed on (111) growth faces of crystals of the cellular iron-storage protein, horse spleen ferritin. Crystal truncation rods (CTR) were measured. A fit of the measured profile of the CTR revealed a surface roughness of 48+/-4.5 A and a top layer spacing contraction of 3.9+/-1.5%. In addition to the peak from the CTR, the rocking curves of the crystals displayed unexpected extra peaks. Multiple scattering is demonstrated to account for them. Future applications of the method could allow the exploration of hydration effects on the growth of protein crystals.

Journal Article↗

Phase sensitive x-ray diffraction imaging of defects in biological macromolecular crystals.

Conventional x-ray diffraction topography is currently used to map defects in the bulk of protein crystals, but the lack of sufficient contrast is frequently a limiting factor. We experimentally demonstrate that this barrier can be circumvented using a method that combines phase sensitive and diffraction imaging principles. Details of defects revealed in tetragonal lysozyme and cubic ferritin crystals are presented and discussed. The approach enabling the detection of the phase changes of diffracted x rays should prove to be useful in the study of defect structures in a broad range of biological macromolecular crystals.

Animals↗

Laboratory multiple-crystal X-ray topography and reciprocal-space mapping of protein crystals: influence of impurities on crystal perfection.

Double-axis multiple-crystal X-ray topography, rocking-curve measurements and triple-axis reciprocal-space mapping have been combined to characterize protein crystals using a laboratory source. Crystals of lysozyme and lysozyme crystals doped with acetylated lysozyme impurities were examined. It was shown that the incorporation of acetylated lysozyme into crystals of lysozyme induces mosaic domains that are responsible for the broadening and/or splitting of rocking curves and diffraction-space maps along the direction normal to the reciprocal-lattice vector, while the overall elastic lattice strain of the impurity-doped crystals does not appear to be appreciable in high angular resolution reciprocal-space maps. Multiple-crystal monochromatic X-ray topography, which is highly sensitive to lattice distortions, was used to reveal the spatial distribution of mosaic domains in crystals which correlates with the diffraction features in reciprocal space. Discussions of the influence of acetylated lysozyme on crystal perfection are given in terms of our observations.

Acetylation↗

Macromolecular impurities and disorder in protein crystals.

The mechanisms by which macromolecular impurities degrade the diffraction properties of protein crystals have been investigated using X-ray topography, high-resolution diffraction line shape measurements, crystallographic data collection, chemical analysis, and two-photon excitation fluorescence microscopy. Hen egg-white lysozyme crystals grown from solutions containing a structurally unrelated protein (ovotransferrin) and a related protein (turkey egg-white lysozyme) can exhibit significantly broadened mosaicity due to formation of cracks and dislocations but have overall B factors and diffraction resolutions comparable to those of crystals grown from uncontaminated lysozyme. Direct fluorescence imaging of the three-dimensional impurity distribution shows that impurities incorporate with different densities in sectors formed by growth on different crystal faces, and that impurity densities in the crystal core and along boundaries between growth sectors can be much larger than in other parts of the crystal. These nonuniformities create stresses that drive formation of the defects responsible for the mosaic broadening. Our results provide a rationale for the use of seeding to obtain high-quality crystals from heavily contaminated solutions and have implications for the use of crystallization for protein purification. Proteins 1999;36:270-281.

Animals↗

Refined structure of orthorhombic lysozyme crystallized at high temperature: correlation between morphology and intermolecular contacts.

The structure of orthorhombic hen egg-white lysozyme (HEWL) crystallized at 310 K has been refined at 1.7 A resolution. Large displacements of the side-chain atoms with respect to the tetragonal structure were observed in many places, in contrast to small displacements of the main-chain atoms. A chloride-ion binding site was observed at an interface of two molecules, but at a different position to the binding site in the tetragonal form. The analysis of intermolecular contacts in the crystal has shown the presence of three independent intermolecular contacts which are called macrobonds A, B and C. Arginine side chains are frequently involved in these macrobonds, suggesting that the high frequency of this residue in HEWL may be a possible reason for the multiple polymorphs of this protein. The crystal forms were determined using a light-reflecting device on a four-circle diffractometer. Correlations between crystal forms and the three-dimensional macrobond networks were interpreted in terms of their components in various crystallographic planes, making use of approximate strengths of hydrogen-bond and van der Waals interatomic forces.

Animals↗

Crystal growth and crystallography.

Selected topics that may be of interest for both crystal-structure and crystal-growth communities are overviewed. The growth of protein crystals, along with that of some other compounds, is one of the topics, and recent insights into related phenomena are considered as examples of applications of general principles. The relationship between crystal growth shape and structure is reviewed and an attempt to introduce semiquantitative characterization of binding for proteins is made. The concept of kinks for complex structures is briefly discussed. Even at sufficiently low supersaturations, the fluctuation of steps may not be sufficient to implement the Gibbs-Thomson law if the kink density is low enough. Subsurface ordering of liquids and growth of rough interfaces from melts is discussed. Crystals growing in microgravity from solution should be more perfect if they preferentially trap stress-inducing impurities, thus creating an impurity-depleted zone around themselves. Evidently, such a zone is developed only around the crystals growing in the absence of convection. Under terrestrial conditions, the self-purified depleted zone is destroyed by convection, the crystal traps more impurity and grows stressed. The stress relief causes mosaicity. In systems containing stress-inducing but poorly trapped impurities, the crystals grown in the absence of convection should be worse than those of their terrestrial counterparts.

Animals↗

Terrestrial and space-grown HAP and OCP crystals: effect of growth conditions on perfection and morphology.

This paper reports comparative characterizations of calcium phosphate crystals grown on earth and in space. At the CaCl2 and KH2PO4 + K2HPO4 solution concentrations and the pH used, only hydroxyapatite (HAP) crystals grow under terrestrial condition while both HAP and octacalcium phosphate (OCP) crystals grew during the space experiment. The space-grown OCP crystals reach 3 mm in size, the space-grown HAP crystals reach sizes up to 100 times larger than the earth-grown crystallites. It was found also that the space-grown crystallites are more perfect than the terrestrial ones, being more stable under electron beam during HRTEM examination. Spherolites of hydroxyapatite consist of small and thin HAP crystals with different orientations. Space-grown OCP crystals containing almost pure OCP phase show strong striations along the c direction due to thickness variations. Terrestrial OCP crystals grown at lowest supersaturation on earth may be almost as large as the space-grown ones, possess a regular habit and are homogeneous in thickness. However, they always contain substantial regions of HAP structure. Also, in these crystals electron irradiation induces phase transformation from crystalline to amorphous (disordered) state during transmission electron microscopy observations. In the space-grown crystals, such transformation needs longer radiation time. We believe that the differences described above come from much lower supersaturation and different pH for crystals nucleating and growing in space compared to those formed on earth.

Calcium Phosphates↗

Calcium phosphate crystallization under terrestrial and microgravity conditions.

Calcium phosphate crystalline powders grown under terrestrial and space (EURECA 1992-1993 flight) conditions in the Solution Growth Facility are analyzed and compared by optical and electron microscopy (scanning and transmission), electron and X-ray microdiffraction and microanalyses. On earth, only small, micrometer size scale, spherolites of hydroxyapatite (HAP) grow. In space, the HAP spherolites reach hundreds of micrometer. Also, octacalcium phosphate (OCP) spherolites up to 3 mm have been obtained. Computer modelling of diffusion in a real chamber has been performed. It suggests high spatial supersaturation gradients at zero gravity which may provide much higher local supersaturations on earth, where convection takes place. The analyses suggest that the dramatic difference between the terrestrial and space samples should come from much lower supersaturation in space.

Calcium Phosphates↗

Crystallization of calcium phosphate in microgravity.

Dilute solutions of CaCl2 and KH2PO4 + K2HPO4 were diffusing from either side into a mixing chamber with KCl solution. The microgravity experiment yielded aggregates of large crystals of OCP (Ca8H2(PO4)3,5H2O) and spherolites of smaller, but still visible crystals of HAP (Ca5OH(PO4)3), the stable final phase. Ground-based experiments yielded submicroscopic HAP crystals. Results of calculations of diffusion and crystal growth on the basis of previous knowledge agree well with observations.

Calcium Phosphates↗