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I Dobrianov

Publications and source records attributed to I Dobrianov.

6 recordsLinked to original sources
PubMed2001

Dynamic response of tetragonal lysozyme crystals to changes in relative humidity: implications for post-growth crystal treatments.

The dynamic response of tetragonal lysozyme crystals to dehydration has been characterized in situ using a combination of X-ray topography, high-resolution diffraction line-shape measurements and conventional crystallographic diffraction. For dehydration from 98% relative humidity (r.h.) to above 89%, mosaicity and diffraction resolution show little change and X-ray topographs remain featureless. Lattice constants decrease rapidly but the lattice-constant distribution within the crystal remains very narrow, indicating that water concentration gradients remain very small. Near 88% r.h., the c-axis lattice parameter decreases abruptly, the steady-state mosaicity and diffraction resolution degrade sharply and topographs develop extensive contrast. This transformation exhibits metastability and hysteresis. At fixed r.h. < 88% it is irreversible, but the original order can be almost completely restored by rehydration. These results suggest that this transformation is a first-order structural transition involving an abrupt loss of crystal water. The front between transformed and untransformed regions may propagate inward from the crystal surface and the resulting stresses along the front may degrade mosaicity. Differences in crystal size, shape and initial perfection may produce the observed variations in degradation timescale. Consequently, the success of more general post-growth treatments may often involve identifying procedures that either avoid lattice transitions, minimize disorder created during such transitions or maintain the lattice in an ordered metastable state.

Crystallography, X-Ray↗
PubMed2000

Enantiomorph determination using inverse reference-beam diffraction images.

It is shown that enantiomorph structures of a noncentrosymmetric crystal can be determined, in the absence of anomalous diffraction signals, by measuring two series of reference-beam oscillation diffraction patterns related by an inverse-beam geometry. The corresponding intensities of the Friedel pairs recorded on the two sets of images exhibit the characteristic three-beam interference effects that provide the unambiguous phase information. The experimental arrangement and the data-analysis procedure are demonstrated through an experimental example on tetragonal lysozyme.

Crystallography, X-Ray↗
PubMed2000

Triplet-phase measurements using reference-beam X-ray diffraction.

Reference-beam diffraction (RBD) is a recently developed phase-sensitive X-ray diffraction technique that incorporates the principle of multiple-beam diffraction into the standard oscillating-crystal data-collection method [Shen (1998). Phys. Rev. Lett. 80, 3268-3271]. Using this technique, a large number of multiple-beam interference profiles can be recorded simultaneously on an area detector, from which a large number of triplet phases of Bragg reflections can be determined in a crystallography experiment. In this article, both the theoretical developments and the experimental procedures of the RBD technique are described in detail. Approximate theoretical approaches for RBD are outlined and simple analytical expressions are obtained that provide the basis for an automated data-analysis procedure that can be used to extract triplet phases from a large number of measured reference-beam diffraction profiles. Experimental examples are given for a variety of crystals including GaAs, tetragonal lysozyme and AlPdMn quasicrystal, using both image plates and a charge-coupled device (CCD) as the area detector. Possible uses of the measured phases for crystal structure determination are discussed as well as future prospects of the RBD technique.

Muramidase↗
PubMed1999

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↗
PubMed1998

X-ray topographic studies of protein crystal perfection and growth.

The effects of solution variations during growth on the perfection of tetragonal lysozyme crystals have been characterized using X-ray topography and high angular and wavevector resolution reciprocal-space scans. X-ray images of crystals grown under nearly uniform conditions show little contrast or evidence of defects, and mosaic widths of these crystals are comparable with those reported for microgravity-grown crystals. Images of crystals for which solution conditions (temperature, pH or salt concentration) are changed after an initial period of uniform growth can show extensive contrast, indicating the presence of disorder. The X-ray mosaic widths of these crystals can be significantly broadened, but their radial widths are at most very slightly broadened, indicating that image contrast is primarily due to mosaicity. Comparison of X-ray images with mosaic scans indicates that regions grown after the change in solution conditions have broader mosaicities and are more disordered; that regions grown immediately after the change tend to have broader mosaicities than subsequent growth regions; and that the pre-change growth region is largely unaffected by solution changes. The observed disorder may arise from solution change-related transient growth instabilities, from transient liquid-liquid phase separation that can occur during the change, and from post-change relaxation of the lattice constant of the pre-change growth regions. These results suggest that solution variations during growth, including those typical of macroseeding, vapor-diffusion growth and other widely used techniques, may be an important source of disorder in some protein crystals.

Animals↗