A double cell for controlling nucleation and growth of protein crystals

1989 ◽  
Vol 22 (2) ◽  
pp. 115-118 ◽  
Author(s):  
M. Przybylska
Keyword(s):  
Crystal Growth ◽  
Protein Crystallization ◽  
Heavy Atom ◽  
Nucleation And Growth ◽  
The Other ◽  
Crystal Nucleation ◽  
Diffusion Method ◽  
Vapour Diffusion ◽  
Reservoir Solution ◽  
The Stability

A simple device for protein crystallization is described that consists of two connected cells, one for the hanging- or sitting-drop vapour diffusion method and the other for changing the concentration of the reservoir solution. It has been found useful for decoupling crystal nucleation from crystal growth, for improving the size and the stability of crystals, and in the preparation of heavy-atom derivatives.

A gradual desiccation method for improving the efficiency of protein crystallization screening

2012 ◽  
Vol 45 (4) ◽  
pp. 758-765 ◽  
Author(s):  
Qin-Qin Lu ◽  
Xu-Zhuo Xie ◽  
Rui-Qing Chen ◽  
Zi-Qing Wu ◽  
Qing-Di Cheng ◽  
...  
Keyword(s):  
Success Rate ◽  
Protein Crystallization ◽  
Diffusion Method ◽  
Powerful Method ◽  
Vapor Diffusion ◽  
Optimal Amount ◽  
Reservoir Solution ◽  
The One

In vapor diffusion protein crystallization screening, it has been reported that replacing the reservoir solution with desiccant can increase the crystallization success rate. Therefore, the desiccation method is a potentially powerful method in practical protein crystallization screening. However, this method is difficult to apply broadly because the optimal amount of desiccant for a specific screening task is unknown. Utilizing an unsuitable amount of desiccant can result in even worse screening results than would be obtained from the traditional vapor diffusion method. Here, it is shown that by employing a modified strategy, named the gradual desiccation method, the problem can be solved without knowing the optimal amount of desiccant, and the crystallization success rate can be further increased compared with the one-time desiccation method.

scholarly journals Peculiarities of Protein Crystal Nucleation and Growth

Crystals ◽  
2018 ◽  
Vol 8 (11) ◽  
pp. 422 ◽  
Author(s):  
Christo Nanev
Keyword(s):  
Crystallization Process ◽  
Protein Crystallization ◽  
Nucleation And Growth ◽  
Crystal Nucleation ◽  
Protein Crystal ◽  
Solution Flow ◽  
Molecular Scale ◽  
Novel Approaches ◽  
Lattice Formation ◽  
Complete Account

This paper reviews investigations on protein crystallization. It aims to present a comprehensive rather than complete account of recent studies and efforts to elucidate the most intimate mechanisms of protein crystal nucleation. It is emphasized that both physical and biochemical factors are at play during this process. Recently-discovered molecular scale pathways for protein crystal nucleation are considered first. The bond selection during protein crystal lattice formation, which is a typical biochemically-conditioned peculiarity of the crystallization process, is revisited. Novel approaches allow us to quantitatively describe some protein crystallization cases. Additional light is shed on the protein crystal nucleation in pores and crevices by employing the so-called EBDE method (equilibration between crystal bond and destructive energies). Also, protein crystal nucleation in solution flow is considered.

The Nature and Crystal Growth of Otoconia in the Rat

1975 ◽  
Vol 84 (1) ◽  
pp. 22-36 ◽  
Author(s):  
Muriel D. Ross ◽  
Donald R. Peacor
Keyword(s):  
Crystal Growth ◽  
Structural Unit ◽  
Integrated Approach ◽  
Nucleation And Growth ◽  
Organic Substrate ◽  
Crystal Nucleation ◽  
Chemical Factor ◽  
Adult Type ◽  
Inorganic Crystal ◽  
Parallel Growth

Several types of otoconia are present in the macular regions of young rats. These include multifaceted, transitional and rounded body forms, some variant otoconia and a few rhombohedrons. The adult form has typically rounded but nonsmooth body surfaces and pointed ends with three planar faces. The multifaceted and transitional otoconia fracture and etch more readily than do the adult type. The differences in properties of the otoconia are considered in the light of known facts concerning inorganic crystal nucleation and growth. This integrated approach indicates that many otoconia originate by seeding of multiple subunits on an organic substrate and develop by the mechanism of parallel growth. The basic structural unit is the rhombohedron. By analogy to inorganic crystals of calcite, it would seem that the typical otoconium grows on the end faces but growth on the side faces is suppressed by some unknown chemical factor. Some otoconia are exceptions, evidently seeding and growing in the pure rhombohedral form. Decalcification of cleaved otoconia shows that organic material is incorporated during growth. The observations are interpreted to indicate that organic substance influences growth and achievement of the adult otoconial form.

scholarly journals Promoting crystallization of antibody–antigen complexesviamicroseed matrix screening

2010 ◽  
Vol 66 (8) ◽  
pp. 927-933 ◽  
Author(s):  
Galina Obmolova ◽  
Thomas J. Malia ◽  
Alexey Teplyakov ◽  
Raymond Sweet ◽  
Gary L. Gilliland
Keyword(s):  
Crystal Growth ◽  
Reliable Method ◽  
Crystal Form ◽  
The Other ◽  
Crystal Nucleation ◽  
Hit Rate ◽  
Fab Fragments ◽  
Crystallization Conditions

The application of microseed matrix screening to the crystallization of antibody–antigen complexes is described for a set of antibodies that include mouse anti-IL-13 antibody C836, its humanized version H2L6 and an affinity-matured variant of H2L6, M1295. The Fab fragments of these antibodies were crystallized in complex with the antigen human IL-13. The initial crystallization screening for each of the three complexes included 192 conditions. Only one hit was observed for H2L6 and none were observed for the other two complexes. Matrix self-microseeding using these microcrystals yielded multiple hits under various conditions that were further optimized to grow diffraction-quality H2L6 crystals. The same H2L6 seeds were also successfully used to promote crystallization of the other two complexes. The M1295 crystals appeared to be isomorphous to those of H2L6, whereas the C836 crystals were in a different crystal form. These results are consistent with the concept that the conditions that are best for crystal growth may be different from those that favor nucleation. Microseed matrix screening using either a self-seeding or cross-seeding approach proved to be a fast, robust and reliable method not only for the refinement of crystallization conditions but also to promote crystal nucleation and increase the hit rate.

scholarly journals The bee Tetragonula builds its comb like a crystal

2020 ◽  
Vol 17 (168) ◽  
pp. 20200187 ◽  
Author(s):  
Silvana S. S. Cardoso ◽  
Julyan H. E. Cartwright ◽  
Antonio G. Checa ◽  
Bruno Escribano ◽  
Antonio J. Osuna-Mascaró ◽  
...  
Keyword(s):  
Crystal Growth ◽  
Lattice Model ◽  
Stingless Bees ◽  
Nucleation And Growth ◽  
Coupled Map Lattice ◽  
Three Dimensions ◽  
Crystal Nucleation ◽  
Target Pattern ◽  
Brood Comb ◽  
Molecular Scale

Stingless bees of the genus Tetragonula construct a brood comb with a spiral or a target pattern architecture in three dimensions. Crystals possess these same patterns on the molecular scale. Here, we show that the same excitable-medium dynamics governs both crystal nucleation and growth and comb construction in Tetragonula , so that a minimal coupled-map lattice model based on crystal growth explains how these bees produce the structures seen in their bee combs.

Examining Protein Crystallization Using Scanning Electron Microscopy

2012 ◽  
Vol 19 (1) ◽  
pp. 145-149 ◽  
Author(s):  
Kathryn Gomery ◽  
Elaine C. Humphrey ◽  
Rodney Herring
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Crystal Growth ◽  
Early Stage ◽  
Protein Crystallization ◽  
Crystal Nucleation ◽  
Protein Crystal ◽  
Microscopy Imaging ◽  
Protein Crystal Growth ◽  
Scanning Electron

AbstractElucidation of protein structure using X-ray crystallography relies on the quality of the crystal. Crystals suffer from many different types of disorder, some of which occur during crystal nucleation and early crystal growth. To date, there are few studies surrounding the quality and nucleation of protein crystals partly due to difficulties surrounding viewing biological samples at high resolution. Recent research has led our current understanding of nucleation to be a two-step mechanism involving the formation of nuclei from dense liquid clusters; it is still unclear whether nuclei first start as amorphous aggregate or as crystalline lattices. Our research examines this mechanism through the use of electron microscopy. Using scanning electron microscopy imaging of the protein crystal growth process, a stacking, spiraling manner of growth is observed. The tops of the pyramid-like tetragonal protein crystal structures measure ~0.2 μm across and contain ~125,000 lysozyme units. This noncrystalline area experiences strain due to growth of the protein crystal. Our work shows that it is possible to view detailed early stage protein crystal growth using a wet scanning electron microscopy technique, thereby overcoming the problem of viewing liquids in a vacuum.

scholarly journals Crystallization and preliminary X-ray diffraction analysis of domain-chimericL-(2S,3S)-butanediol dehydrogenase

2014 ◽  
Vol 70 (4) ◽  
pp. 461-463
Author(s):  
Tomohito Shimegi ◽  
Takuji Ooyama ◽  
Takashi Ohtsuki ◽  
Genji Kurisu ◽  
Masami Kusunoki ◽  
...  
Keyword(s):  
Synchrotron Radiation ◽  
Diffusion Method ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Hanging Drop ◽  
X Ray ◽  
Enzymatic Function ◽  
Vapour Diffusion ◽  
The Stability ◽  
Butanediol Dehydrogenase

A domain-chimeric L-2,3-butanediol dehydrogenase (chimera L-BDH), which was designed to possess both theS-configuration specificity of L-BDH and the stability ofmeso-BDH, was constructed by exchanging the respective domains of these two BDHs. However, chimera L-BDH possessed a lower enzymatic function than expected based on the two original enzymes. To elucidate the causes of the decreased stability and substrate specificity, crystallization of the protein was performed. Chimera L-BDH was purified to homogeneityviaammonium sulfate fractionation and three column-chromatography steps, and was crystallized using the hanging-drop vapour-diffusion method. The crystals belonged to space groupC2221, diffracted synchrotron radiation to 1.58 Å resolution and were most likely to contain two molecules in the asymmetric unit.

Comparative Studies of Protein Crystallization by Vapour-Diffusion and Microbatch Techniques

1998 ◽  
Vol 54 (1) ◽  
pp. 8-15 ◽  
Author(s):  
Naomi E. Chayen
Keyword(s):  
Comparative Studies ◽  
Protein Crystallization ◽  
The Other ◽  
Topical Review ◽  
Vapour Diffusion ◽  
Crystallization Conditions

Numerous reports have been published in the literature which describe the crystallization of macromolecules by a variety of crystallization methods, including the vapour-diffusion and microbatch techniques. This topical review compares the results of examples of proteins which were crystallized by both vapour-diffusion and microbatch methods. The inherent features of the vapour-diffusion and microbatch methods are discussed and some specific conditions where one method appears more favourable than the other are reported. Guidelines for the conversion of crystallization conditions from vapour diffusion to microbatch (and vice versa) are also presented.

Use of super-water-repellent fractal surfaces in the crystallization of macromolecules

1999 ◽  
Vol 55 (6) ◽  
pp. 1247-1249 ◽  
Author(s):  
Isao Fujii ◽  
Noriaki Hirayama
Keyword(s):  
Protein Crystallization ◽  
Diffusion Method ◽  
Water Repellent ◽  
Hen Egg White Lysozyme ◽  
Fractal Surface ◽  
Fractal Surfaces ◽  
Egg White Lysozyme ◽  
Crystallization Experiments ◽  
Vapour Diffusion ◽  
Hen Egg White

The effect of fractal surfaces on protein crystallization has been investigated. A super-water-repellent fractal surface consisting of alkylketene dimers was formed on microscope cover slips. Crystallization experiments with hen egg-white lysozyme using the sitting-drop vapour-diffusion method have been undertaken on the fractal and conventionally siliconized surfaces. The size of the crystals obtained on the fractal surface is significantly larger than on the conventionally siliconized surface. The number of crystals on the fractal surface is markedly smaller than on the conventionally siliconized surface. The results clearly indicate that the fractal surface is very effective and promising in the crystallization of macromolecules using the sitting-drop vapour-diffusion method.

Uniformity of Magnification from Different Electron Microscopes

1967 ◽  
Vol 25 ◽  
pp. 32-33
Author(s):  
Godfrey C. Hoskins ◽  
V. Williams ◽  
V. Allison
Keyword(s):  
Diffraction Grating ◽  
The Other ◽  
Carbon Replica ◽  
Electron Microscopes ◽  
The Stability

The method demonstrated is an adaptation of a proven procedure for accurately determining the magnification of light photomicrographs. Because of the stability of modern electrical lenses, the method is shown to be directly applicable for providing precise reproducibility of magnification in various models of electron microscopes.A readily recognizable area of a carbon replica of a crossed-line diffraction grating is used as a standard. The same area of the standard was photographed in Phillips EM 200, Hitachi HU-11B2, and RCA EMU 3F electron microscopes at taps representative of the range of magnification of each. Negatives from one microscope were selected as guides and printed at convenient magnifications; then negatives from each of the other microscopes were projected to register with these prints. By deferring measurement to the print rather than comparing negatives, correspondence of magnification of the specimen in the three microscopes could be brought to within 2%.

Sign in / Sign up

Export Citation Format

Share Document