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Hanging-drop Vapour Diffusion (hanging-drop + vapour_diffusion)
Selected AbstractsCrystallization and preliminary X-ray analysis of RecG, a replication-fork reversal helicase from Thermotoga maritima complexed with a three-way DNA junctionACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2001Martin R. Singleton The monomeric 3,-5, helicase RecG from the thermophilic bacterium Thermotoga maritima has been crystallized in complex with a three-way DNA junction, the preferred physiological substrate. The crystals were obtained by hanging-drop vapour diffusion. The crystals belong to space group C2, with unit-cell parameters a = 133.7, b = 144.6, c = 84.0,Å, , = 113.8°. Native data to a resolution of 3.25,Å were collected from crystals flash-cooled to 100,K. [source] Crystallization and preliminary X-ray diffraction analysis of rat autotaxinACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2010Jacqueline E. Day Rat autotaxin has been cloned, expressed, purified to homogeneity and crystallized via hanging-drop vapour diffusion using PEG 3350 as precipitant and ammonium iodide and sodium thiocyanate as salts. The crystals diffracted to a maximum resolution of 2.05,Å and belonged to space group P1, with unit-cell parameters a = 53.8, b = 63.3, c = 70.5,Å, , = 98.8, , = 106.2, , = 99.8°. Preliminary X-ray diffraction analysis indicated the presence of one molecule per asymmetric unit, with a solvent content of 47%. [source] Crystallization and preliminary crystallographic analysis of the ADP-ribosyltransferase HopU1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 8 2010Yan Lin Several Gram-negative pathogens of plants and animals and some eukaryotic associated bacteria use type III protein-secretion systems (T3SSs) to deliver bacterial virulence-associated `effector' proteins directly into host cells. HopU1 is a type III effector protein from the plant pathogen Pseudomonas syringae, which causes plant bacterial speck disease. HopU1 quells host immunity through ADP-ribosylation of GRP7 as a substrate. HopU1 has been reported as the first ADP-ribosyltransferase virulence protein to be identified in a plant pathogen. Although several structures of ADP-ribosyltransferases have been determined to date, no structure of an ADP-ribosyltransferase from a plant pathogen has been determined. Here, the protein expression, purification, crystallization and preliminary crystallographic analysis of HopU1 are reported. Diffracting crystals were grown by hanging-drop vapour diffusion using polyethylene glycol 10,000 as a precipitant. Native and SAD data sets were collected using native and selenomethionine-derivative HopU1 crystals. The diffraction pattern of the crystal extended to 2.7,Å resolution using synchrotron radiation. The crystals belonged to space group P43, with unit-cell parameters a = 92.6, b = 92.6, c = 101.6,Å. [source] Purification, crystallization and preliminary X-ray analysis of the PCNA2,PCNA3 complex from Sulfolobus tokodaii strain 7ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2009Akito Kawai Crenarchaeal PCNA is known to consist of three subunits (PCNA1, PCNA2 and PCNA3) that form a heterotrimer (PCNA123). Recently, another heterotrimeric PCNA composed of only PCNA2 and PCNA3 was identified in Sulfolobus tokodaii strain 7 (stoPCNAs). In this study, the purified stoPCNA2,stoPCNA3 complex was crystallized by hanging-drop vapour diffusion. The crystals obtained belonged to the orthorhombic space groups I222 and P21212, with unit-cell parameters a = 91.1, b = 111.8, c = 170.9,Å and a = 91.1, b = 160.6, c = 116.6,Å, respectively. X-ray diffraction data sets were collected to 2.90,Å resolution for the I222 crystals and to 2.80,Å resolution for the P21212 crystals. [source] Crystallization and preliminary X-ray diffraction analysis of the lectin from Canavalia boliviana Piper seedsACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2009Tales Rocha Moura Plant lectins are the most studied group of carbohydrate-binding proteins. Despite the high similarity between the members of the Diocleinae subtribe (Leguminosae) group, they present differing biological activities. Canavalia boliviana lectin (Cbol) was purified using a Sephadex G-50 column and crystallized in the presence of X-Man by hanging-drop vapour diffusion at 293,K. After optimization, crystals suitable for diffraction were obtained under the condition 0.1,M HEPES pH 7.5 and 3.0,M sodium formate. The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 126.70, b = 66.64, c = 64.99,Å, , = 90.0, , = 120.8, , = 90.0°. Assuming the presence of a dimer in the asymmetric unit, the solvent content was estimated to be about 46%. A complete data set was collected at 1.5,Å resolution. [source] Crystallization and preliminary crystallographic studies of the recombinant l - N -carbamoylase from Geobacillus stearothermophilus CECT43ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2008Sergio Martínez-Rodríguez N -Carbamoyl- l -amino-acid amidohydrolases (l - N -carbamoylases; EC 3.5.1.87) hydrolyze the carbon,nitrogen bond of the ureido group in N -carbamoyl- l -,-amino acids. These enzymes are commonly used in the production of optically pure natural and non-natural l -amino acids using the `hydantoinase process'. Recombinant l - N -carbamoylase from Geobacillus stearothermophilus CECT43 has been expressed, purified and crystallized by hanging-drop vapour diffusion. X-ray data were collected to a resolution of 2.75,Å. The crystals belonged to space group P21212, with unit-cell parameters a = 103.2, b = 211.7, c = 43.1,Å and two subunits in the asymmetric unit. [source] Incorporation of a disaccharide nucleoside into the backbone of double-stranded DNA: crystallization and preliminary X-ray diffractionACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 10 2005Kristof Van Hecke Incorporation of a disaccharide nucleoside into double-stranded DNA can be considered as a chemical (non-enzymatic) alternative for site-specific cleavage of DNA. Crystals of the sequence d(CGCGAATT*CGCG), where * is an incorporated ribose, were obtained by hanging-drop vapour diffusion and diffracted to 2.6,Å. The crystals belong to the orthorhombic space group P2221, with unit-cell parameters a = 41.52, b = 57.63, c = 81.39,Å, indicating a new crystal packing motif for an oligonucleotide dodecamer sequence. [source] | ||||||||||