Home About us Contact
|
Home About us Contact | ||
|
|
||
Sitting-drop Vapour-diffusion Method (sitting-drop + vapour-diffusion_method)
Selected AbstractsCrystallization and preliminary X-ray analysis of cellobiose phosphorylase from Cellvibrio gilvusACTA CRYSTALLOGRAPHICA SECTION D, Issue 10 2004Masafumi Hidaka A recombinant cellobiose phosphorylase from Cellvibrio gilvus has been prepared and crystallized by the sitting-drop vapour-diffusion method using 10,mg,ml,1 purified enzyme, 1.5,M ammonium sulfate, 0.1,M MES buffer pH 7.0 and 5,mM glucose. A suitable crystal was obtained after 10,d incubation at 298,K. The crystal belongs to space group P21, with unit-cell parameters a = 84.77, b = 98.31, c = 104.04,Å, , = 102.73°. X-ray diffraction data to 2.1,Å resolution have been collected at KEK-PF BL-5A. [source] Crystallization and preliminary X-ray diffraction study of mammalian mitochondrial seryl-tRNA synthetaseACTA CRYSTALLOGRAPHICA SECTION D, Issue 7 2004Sarin Chimnaronk The mitochondrial seryl-tRNA synthetase (mt SerRS) from Bos taurus was overexpressed in Escherichia coli and crystallized using the sitting-drop vapour-diffusion method. Crystals grew in a very narrow range of conditions using PEG 8000 as precipitant at room temperature. An appropriate concentration of lithium sulfate was critical for crystal nucleation. Crystals diffracted well beyond a resolution of 1.6,Å and were found to belong to the orthorhombic space group C2221, with unit-cell parameters a = 79.89, b = 230.42, c = 135.60,Å. There is one dimer (Mr, 113,kDa) in the asymmetric unit, with a solvent content of 55%. Efforts to solve the phase problem by molecular replacement are under way. [source] Purification, crystallization and preliminary X-ray diffraction analysis of human oncoprotein SET/TAF-1,ACTA CRYSTALLOGRAPHICA SECTION D, Issue 4 2004Shinsuke Muto The human oncoprotein SET/TAF-1, has been crystallized by the sitting-drop vapour-diffusion method using ammonium sulfate as a precipitant. The crystal belongs to space group C2, with unit-cell parameters a = 119.6, b = 62.8, c = 61.0,Å, , = 89.7°, and contains two molecules in the asymmetric unit. A complete data set was collected to 2.8,Å resolution using synchrotron radiation. [source] Crystallization and preliminary X-ray studies of xylanase 10B from Thermotoga maritimaACTA CRYSTALLOGRAPHICA SECTION D, Issue 9 2003Ihsanawati Xylanases catalyze the hydrolysis of the ,-1,4-glycosidic bonds of xylan, which is the second most abundant component of plant cell walls after cellulose. The recombinant xylanase 10B from Thermotoga maritima MSB8 was prepared and crystallized by the sitting-drop vapour-diffusion method using 40,mM zinc acetate, 20,mM MES buffer pH 6.0 and 3% ethanol. Intensity data were collected to 2.5,Å resolution at beamline BL26B2 of SPring-8. Preliminary X-ray analysis showed that the crystal belongs to space group P21212, with unit-cell parameters a = 77.3, b = 80.6, c = 58.2,Å and one molecule per asymmetric unit. [source] Crystallization and preliminary X-ray analysis of Yersinia pseudotuberculosis -derived mitogenACTA CRYSTALLOGRAPHICA SECTION D, Issue 7 2003Roberta Donadini Yersinia pseudotuberculosis -derived mitogen (YPM), a superantigen with no amino-acid sequence similarity to other known superantigens, has been crystallized by the sitting-drop vapour-diffusion method. The crystals belong to space group C2, with unit-cell parameters a = 138.67, b = 78.66, c = 32.91,Å, , = 91.97°. A native data set has been collected to a resolution of 1.8,Å using synchrotron radiation. Self-rotation function calculations suggest the presence of three molecules in the asymmetric unit, corresponding to a solvent content of 45%. [source] Crystallization and preliminary X-ray diffraction studies of a novel alcohol dehydrogenase from the hyperthermophilic archaeon Aeropyrum pernixACTA CRYSTALLOGRAPHICA SECTION D, Issue 1 2003Jodie E. Guy A novel alcohol dehydrogenase enzyme has been cloned from the hyperthermophilic archaeon Aeropyrum pernix and overexpressed in Escherichia coli. This zinc-containing enzyme has been crystallized by the sitting-drop vapour-diffusion method using PEG 600 as precipitant. The crystals diffract to 1.5,Å resolution and belong to the orthorhombic space group P21212, with unit-cell parameters a = 100.7, b = 103.2, c = 67.5,Å. The asymmetric unit contains two enzyme monomers. Two synchrotron data sets have been collected: one at a wavelength near the absorption edge of zinc and one at a remote wavelength. Three strong zinc-ion positions were visible in the anomalous Patterson map. Two additional weaker zinc ions have been identified by anomalous Fourier synthesis. [source] Structure of cytochrome c2 from Rhodospirillum centenumACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2001A. Camara-Artigas Cytochrome c2 from the purple photosynthetic bacterium Rhodospirillum centenum has been crystallized by the sitting-drop vapour-diffusion method. The crystals belong to the orthorhombic space group P212121, with unit-cell parameters a = 29.7, b = 59.9, c = 65.4,Å, and diffract to a resolution limit of 1.7,Å. The Fe-atom position was determined from its anomalous scattering contribution and a molecular-replacement solution was calculated. The correctness of the solution was confirmed by parallel isomorphous replacement studies. The resulting model has a type I cytochrome fold with two features, an extended ,-helix and a surface-charge distribution, that are distinctive to this protein. The implications of these structural features for the ability of the cytochrome to serve as an electron carrier are discussed. [source] Crystallization and preliminary X-ray diffraction studies of a novel alkaline serine protease (KP-43) from alkaliphilic Bacillus sp. strain KSM-KP43ACTA CRYSTALLOGRAPHICA SECTION D, Issue 5 2001Tsuyoshi Nonaka A novel alkaline serine protease (KP-43) which belongs to a new class of the subtilisin superfamily was crystallized by the sitting-drop vapour-diffusion method with ammonium sulfate as a precipitant. The crystals belong to the orthorhombic space group C2221, with unit-cell parameters a = 43.50,(2), b = 110.4,(1), c = 168.9,(1),Å. The crystals diffract X-rays beyond 1.9,Å resolution using Cu,K, radiation from a rotating-anode generator and are suitable for high-resolution crystal structure analysis. [source] Crystallization and preliminary X-ray diffraction analysis of a rat biliverdin reductaseACTA CRYSTALLOGRAPHICA SECTION D, Issue 9 2000Danyu Sun Biliverdin reductase (BVR) catalyzes the final step of haem degradation and converts biliverdin to bilirubin using NAD(P)H as an electron donor. This paper deals with the first crystallization and preliminary crystallographic study of recombinant rat BVR expressed in Escherichia coli. Crystals of BVR were obtained by the sitting-drop vapour-diffusion method. Using synchrotron radiation at station BL44B2 of SPring-8, Japan, BVR diffraction data were collected to 1.6,Å resolution. Crystals belong to the orthorhombic space group P212121, with unit-cell parameters a = 58.89, b = 70.41, c = 87.76,Å. The complete determination of the crystallographic structure is currently in progress using MAD (multiwavelength anomalous diffraction) data from an Ir-derivative crystal. [source] Crystallization and preliminary crystallographic characterization of the iron-regulated outer membrane lipoprotein FrpD from Neisseria meningitidisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2010Ekaterina Sviridova Fe-regulated protein D (FrpD) is a Neisseria meningitidis outer membrane lipoprotein that may be involved in the anchoring of the secreted repeat in toxins (RTX) protein FrpC to the outer bacterial membrane. However, the function and biological roles of the FrpD and FrpC proteins remain unknown. Native and selenomethionine-substituted variants of recombinant FrpD43,271 protein were crystallized using the sitting-drop vapour-diffusion method. Diffraction data were collected to a resolution of 2.25,Å for native FrpD43,271 protein and to a resolution of 2.00,Å for selenomethionine-substituted FrpD43,271 (SeMet FrpD43,271) protein. The crystals of native FrpD43,271 protein belonged to the hexagonal space group P62 or P64, while the crystals of SeMet FrpD43,271 protein belonged to the primitive orthorhombic space group P212121. [source] Overexpression, crystallization and preliminary X-ray analysis of xylulose-5-phosphate/fructose-6-phosphate phosphoketolase from Bifidobacterium breveACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 8 2010Ryuichiro Suzuki The xylulose-5-phosphate/fructose-6-phosphate phosphoketolase gene from Bifidobacterium breve was cloned and overexpressed in Escherichia coli. The enzyme was purified to homogeneity and crystallized by the sitting-drop vapour-diffusion method. Crystals were obtained at 293,K using 0.05,mM thiamine diphosphate, 0.25,mM MgCl2, 24%(w/v) PEG 6000 and 0.1,M Bicine pH 9.0. The crystals belonged to the tetragonal space group I422, with unit-cell parameters a = b = 174.8, c = 163.8,Å, and diffracted to beyond 1.7,Å resolution. [source] Expression, purification and crystallization of a plant type III polyketide synthase that produces diarylheptanoidsACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 8 2010Hiroyuki Morita Curcuminoid synthase (CUS) from Oryza sativa is a plant-specific type III polyketide synthase that catalyzes the one-pot formation of bisdemethoxycurcumin by the condensation of two molecules of 4-coumaroyl-CoA and one molecule of malonyl-CoA. Recombinant CUS was expressed in Escherichia coli and crystallized by the sitting-drop vapour-diffusion method. The crystals belonged to space group P21, with unit-cell parameters a = 72.7, b = 97.2, c = 126.2,Å, , = , = 90.0, , = 103.7°. A diffraction data set was collected in-house to 2.5,Å resolution. [source] Crystallization and preliminary X-ray analysis of tubulin-folding cofactor A from Arabidopsis thalianaACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 8 2010Lu Lu Tubulin-folding cofactor A (TFC A) is a molecular post-chaperonin that is involved in the ,-tubulin-folding pathway. It has been identified in many organisms including yeasts, humans and plants. In this work, Arabidopsis thaliana TFC A was expressed in Escherichia coli and purified to homogeneity. After thrombin cleavage, a well diffracting crystal was obtained by the sitting-drop vapour-diffusion method at 289,K. The crystal diffracted to 1.6,Å resolution using synchrotron radiation and belonged to space group I41, with unit-cell parameters a = 55.0, b = 55.0, c = 67.4,Å. [source] Structure of the newly found green turtle egg-white ribonucleaseACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2010Somporn Katekaew Marine green turtle (Chelonia mydas) egg-white ribonuclease (GTRNase) was crystallized from 1.1,M ammonium sulfate pH 5.5 and 30% glycerol using the sitting-drop vapour-diffusion method. The structure of GTRNase has been solved at 1.60,Å resolution by the molecular-replacement technique using a model based on the structure of RNase 5 (murine angiogenin) from Mus musculus (46% identity). The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 86.271, b = 34.174, c = 39.738,Å, , = 90, , = 102, , = 90°. GTRNase consists of three helices and seven ,-strands and displays the ,+, folding topology typical of a member of the RNase A superfamily. Superposition of the C, coordinates of GTRNase and RNase A superfamily members indicates that the overall structure is highly similar to that of angiogenin or RNase 5 from M. musculus (PDB code 2bwl) and RNase A from Bos taurus (PDB code 2blz), with root-mean-square deviations of 3.9 and 2.0,Å, respectively. The catalytic residues are conserved with respect to the RNase A superfamily. The three disulfide bridges observed in the reptilian enzymes are conserved in GTRNase, while one further disulfide bond is required for the structural stability of mammalian RNases. GTRNase is expressed in egg white and the fact that its sequence has the highest similarity to that of snapping turtle pancreatic RNase suggests that the GTRNase secreted from oviduct cells to form egg white is probably the product of the same gene as activated in pancreatic cells. [source] Expression, crystallization and preliminary X-ray crystallographic study of ethanolamine ammonia-lyase from Escherichia coliACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 6 2010Naoki Shibata Ethanolamine ammonia-lyase (EAL) catalyzes the adenosylcobalamin-dependent conversion of ethanolamine to acetaldehyde and ammonia. The wild-type enzyme shows a very low solubility. N-terminal truncation of the Escherichia coli EAL ,-subunit dramatically increases the solubility of the enzyme without altering its catalytic properties. Two deletion mutants of the enzyme [EAL(,,4,30) and EAL(,,4,43)] have been overexpressed, purified and crystallized using the sitting-drop vapour-diffusion method. Crystals of EAL(,,4,30) and EAL(,,4,43) diffracted to approximately 8.0 and 2.1,Å resolution, respectively. [source] Crystallization and preliminary X-ray analysis of SDR-type pyridoxal dehydrogenase from Mesorhizobium lotiACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 6 2010Huy Nhat Chu Pyridoxal 4-dehydrogenase from Mesorhizobium loti MAFF303099 was overexpressed in Escherichia coli. The recombinant selenomethionine-substituted enzyme was purified and crystallized by the sitting-drop vapour-diffusion method using PEG 4000 as precipitant. Crystals grew in the presence of 0.45,mM NAD+. The crystals diffracted to 2.9,Å resolution and belonged to the monoclinic space group P21, with unit-cell parameters a = 86.20, b = 51.11, c = 91.73,Å, , = 89.36°. The calculated VM values suggested that the asymmetric unit contained four molecules. [source] Crystallization and preliminary X-ray crystallographic analysis of a full-length active form of the Cry4Ba toxin from Bacillus thuringiensisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 6 2010Niramon Thamwiriyasati To obtain a complete structure of the Bacillus thuringiensis Cry4Ba mosquito-larvicidal protein, a 65,kDa functional form of the Cry4Ba-R203Q mutant toxin was generated for crystallization by eliminating the tryptic cleavage site at Arg203. The 65,kDa trypsin-resistant fragment was purified and crystallized using the sitting-drop vapour-diffusion method. The crystals belonged to the rhombohedral space group R32, with unit-cell parameters a = b = 184.62, c = 187.36,Å. Diffraction data were collected to at least 2.07,Å resolution using synchrotron radiation and gave a data set with an overall Rmerge of 9.1% and a completeness of 99.9%. Preliminary analysis indicated that the asymmetric unit contained one molecule of the active full-length mutant, with a VM coefficient and solvent content of 4.33,Å3,Da,1 and 71%, respectively. [source] Cloning, overexpression, purification, crystallization and preliminary X-ray analysis of a female-specific lipocalin (FLP) expressed in the lacrimal glands of Syrian hamstersACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2010Ved Prakash Dubey Proteins belonging to the lipocalin superfamily are usually secretory proteins of molecular mass ,20,kDa with a hydrophobic pocket for the binding and transport of diverse small ligands. Various lipocalins have been associated with many biological processes, e.g. immunomodulation, odorant transport, pheromonal activity, retinoid transport, cancer-cell interactions etc. However, the exact functions of many lipocalins and the ligands bound by them are unclear. Previously, the cDNA of a 20,kDa lipocalin (FLP) which is female-specifically expressed in the lacrimal glands of Syrian (golden) hamsters and secreted in the tears of females has been identified and cloned. His-tagged recombinant FLP (rFLP) has now been cloned, overexpressed in Escherichia coli as a soluble protein and purified to homogeneity using Ni-affinity followed by size-exclusion chromatography. Purified rFLP was crystallized using the sitting-drop vapour-diffusion method. The crystals tested belonged to space group P212121 and diffracted to beyond 1.86,Å resolution. Solvent-content analysis indicated the presence of one monomer in the asymmetric unit. [source] Preliminary X-ray crystallographic analysis of SMU.2055 protein from the caries pathogen Streptococcus mutansACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2010Wang-Hong Zhao The SMU.2055 gene from the major caries pathogen Streptococcus mutans is annotated as a putative acetyltransferase with 163 amino-acid residues. In order to identify its function via structural studies, the SMU.2055 gene was cloned into the expression vector pET28a. Native and SeMet-labelled SMU.2055 proteins with a His6 tag at the N-terminus were expressed at a high level in Escherichia coli strain BL21 (DE3) and purified to homogeneity by Ni2+ -chelating affinity chromatography. Diffraction-quality crystals of SeMet-labelled SMU.2055 were obtained using the sitting-drop vapour-diffusion method and diffracted to a resolution of 2.5,Å on beamline BL17A at the Photon Factory, Tsukuba, Japan. The crystals belong to the orthorhombic space group C2221, with unit-cell parameters a = 92.0, b = 95.0, c = 192.2,Å. The asymmetric unit contained four molecules, with a solvent content of 57.1%. [source] Crystallization and preliminary crystallographic analysis of the central domain of Drosophila Dribble, a protein that is essential for ribosome biogenesisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2010Tat-Cheung Cheng Dribble (DBE) is a Drosophila protein that is essential for ribosome biogenesis. Bioinformatics analysis revealed a folded central domain of DBE which is flanked by structural disorder in the N- and C-terminal regions. The protein fragment spanning amino-acid residues 16,197 (DBE16,197) was produced for structural determination. In this report, the crystallization and preliminary X-ray diffraction data analysis of the DBE16,197 protein domain are described. Crystals of DBE16,197 were grown by the sitting-drop vapour-diffusion method at 289,K using ammonium phosphate as a precipitant. The crystals belonged to space group P212121. Data were collected that extended to beyond 2,Å resolution. [source] Crystallization and preliminary X-ray analysis of the diadenosine 5,,5,,,- P1,P4 -tetraphosphate phosphorylase from Mycobacterium tuberculosis H37RvACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2010Shigetarou Mori A novel diadenosine 5,,5,,,- P1,P4 -tetraphosphate (Ap4A) phosphorylase (Rv2613c) from Mycobacterium tuberculosis H37Rv has been crystallized by the sitting-drop vapour-diffusion method. The crystal belonged to space group C2, with unit-cell parameters a = 101.5, b = 63.6, c = 79.1,Å, , = 110.9°. The diffraction of the crystals extended to 1.9,Å resolution. The asymmetric unit is expected to contain two molecules of Rv2613c, with a corresponding crystal volume per protein weight (VM) of 2.41,Å3,Da,1 and a solvent content of 49.1%. This is the first report of a crystal of Ap4A phosphorylase. [source] Crystallization and preliminary crystallographic analysis of maganese(II)-dependent 2,3-dihydroxybiphenyl 1,2-dioxygenase from Bacillus sp.ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2010A thermostable manganese(II)-dependent 2,3-dihydroxybiphenyl-1,2-dioxygenase derived from Bacillus sp. JF8 was crystallized. The initial screening for crystallization was performed by the sitting-drop vapour-diffusion method using a crystallization robot, resulting in the growth of two crystal forms. The first crystal belonged to space group P1, with unit-cell parameters a = 62.7, b = 71.4, c = 93.6,Å, , = 71.2, , = 81.0, , = 64.0°, and diffracted to 1.3,Å resolution. The second crystal belonged to space group I222, with unit-cell parameters a = 74.2, b = 90.8, c = 104.3,Å, and diffracted to 1.3,Å resolution. Molecular-replacement trials using homoprotocatechuate 2,3-dioxygenase from Arthrobacter globiformis (28% amino-acid sequence identity) as a search model provided a satisfactory solution for both crystal forms. [source] Overproduction, purification, crystallization and preliminary X-ray diffraction analysis of Trypanosoma brucei gambiense glycerol kinaseACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2010Emmanuel Oluwadare Balogun In the bloodstream forms of human trypanosomes, glycerol kinase (GK; EC 2.7.1.30) is one of the nine glycosomally compartmentalized enzymes that are essential for energy metabolism. In this study, a recombinant Trypanosoma brucei gambiense GK (rTbgGK) with an N-terminal cleavable His6 tag was overexpressed, purified to homogeneity and crystallized by the sitting-drop vapour-diffusion method using PEG 400 as a precipitant. A complete X-ray diffraction data set to 2.75,Å resolution indicated that the crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 63.84, b = 121.50, c = 154.59,Å. The presence of two rTbgGK molecules in the asymmetric unit gives a Matthews coefficient (VM) of 2.5,Å3,Da,1, corresponding to 50% solvent content. [source] Crystallization and preliminary crystallographic analysis of mouse peroxiredoxin II with significant pseudosymmetryACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2010Ari Ora Peroxiredoxin II was cloned from mouse B cells into pCold 1 expression vector and produced as a His-tagged recombinant protein in Escherichia coli. A ring form was isolated by gel filtration. A crystal obtained by the sitting-drop vapour-diffusion method diffracted to 1.77,Å resolution at 100,K. The crystal belonged to space group P21212, with unit-cell parameters a = 117.4, b = 133.9, c = 139.1,Å. The asymmetric unit is expected to contain six dimers of peroxiredoxin II, with a corresponding solvent content of 39.3%. Peaks in the native Patterson function together with pseudo-systematic absences suggested that the crystals suffered from severe translational pseudosymmetry. [source] Purification, crystallization and preliminary crystallographic analysis of the minor pilin FctB from Streptococcus pyogenesACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 2 2010Christian Linke The minor pilin FctB is an integral part of the pilus assembly expressed by Streptococcus pyogenes. Since it is located at the cell wall, it can be hypothesized that it functions as a cell-wall anchor for the streptococcal pilus. In order to elucidate its structure, the genes for FctB from the S. pyogenes strains 90/306S and SF370 were cloned for overexpression in Escherichia coli. FctB from strain 90/306S was crystallized by the sitting-drop vapour-diffusion method using sodium citrate as a precipitant. The hexagonal FctB crystals belonged to space group P61 or P65, with unit-cell parameters a = b = 95.15, c = 100.25,Å, and diffracted to 2.9,Å resolution. [source] Crystallization and preliminary crystallographic analysis of recombinant VSP1 from Arabidopsis thalianaACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 2 2010Zhu-Bing Shi VSP1 is a defence protein in Arabidopsis thaliana that may also be involved in control of plant development. The recombinant protein has been overexpressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method. The crystal diffracted to 1.9,Å resolution and a complete X-ray data set was collected at 100,K using Cu,K, radiation from a rotating-anode X-ray source. The crystals belonged to space group C2. As there are no related structures that could be used as a search model for molecular replacement, work is in progress on experimental phasing using heavy-atom derivatives and selenomethionine derivatives. [source] Crystallization and preliminary X-ray analysis of NADH:rubredoxin oxidoreductase from Clostridium acetobutylicumACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 1 2010Koji Nishikawa NADH:rubredoxin oxidoreductase (NROR), an O2 -inducible protein, is a versatile electron donor for scavengers of O2 and reactive oxygen species (ROS) in Clostridium acetobutylicum. Recombinant NROR was overexpressed in Escherichia coli and purified to homogeneity; it was subsequently crystallized using the sitting-drop vapour-diffusion method at 293,K. Preliminary crystallographic analysis revealed that the crystals belonged to space group P4122 or P4322, with unit-cell parameters a = b = 98.6, c = 88.3,Å, and diffracted to 2.1,Å resolution. Assuming that the crystals contained one molecule per asymmetric unit, the Matthews coefficient was calculated to be 2.7,Å3,Da,1 and the solvent content to be 54.1%. [source] Crystallization and preliminary X-ray diffraction analysis of the P3 RNA domain of yeast ribonuclease MRP in a complex with RNase P/MRP protein components Pop6 and Pop7ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 1 2010Anna Perederina Eukaryotic ribonucleases P and MRP are closely related RNA-based enzymes which contain a catalytic RNA component and several protein subunits. The roles of the protein subunits in the structure and function of eukaryotic ribonucleases P and MRP are not clear. Crystals of a complex that included a circularly permuted 46-nucleotide-long P3 domain of the RNA component of Saccharomyces cerevisiae ribonuclease MRP and selenomethionine derivatives of the shared ribonuclease P/MRP protein components Pop6 (18.2,kDa) and Pop7 (15.8,kDa) were obtained using the sitting-drop vapour-diffusion method. The crystals belonged to space group P4222 (unit-cell parameters a = b = 127.2, c = 76.8,Å, , = , = , = 90°) and diffracted to 3.25,Å resolution. [source] Crystallization and preliminary X-ray analysis of a monomeric mutant of Azami-Green (mAG), an Aequorea victoria green fluorescent protein-like green-emitting fluorescent protein from the stony coral Galaxea fascicularisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2009Tatsuki Ebisawa Monomeric Azami-Green (mAG) from the stony coral Galaxea fascicularis is the first monomeric green-emitting fluorescent protein that is not a derivative of Aequorea victoria green fluorescent protein (avGFP). mAG and avGFP are 27% identical in amino-acid sequence. Diffraction-quality crystals of recombinant mAG were obtained by the sitting-drop vapour-diffusion method using PEG 3350 as the precipitant. The mAG crystal diffracted X-rays to 2.20,Å resolution on beamline AR-NW12A at the Photon Factory (Tsukuba, Japan). The crystal belonged to space group P1, with unit-cell parameters a = 41.78, b = 51.72, c = 52.89,Å, , = 90.96, , = 103.41, , = 101.79°. The Matthews coefficient (VM = 2.10,Å3,Da,1) indicated that the crystal contained two mAG molecules per asymmetric unit. [source] Crystallization and preliminary X-ray crystallographic analysis of blood coagulation factor V-activating proteinase (RVV-V) from Russell's viper venomACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 12 2009Daisuke Nakayama Russell's viper venom blood coagulation factor V activator (RVV-V) is a thrombin-like serine proteinase that specifically activates factor V by cleaving a single peptide bond between Arg1545 and Ser1546. Activated factor V combines with activated factor X produced by the enzyme RVV-X in the venom to form the prothombinase complex, which can induce disseminated intravascular coagulopathy in envenomated animals. In the current study, RVV-V was crystallized in order to attempt to understand its substrate specificity for factor V. Four distinct crystal forms of RVV-V were obtained using the sitting-drop vapour-diffusion method and diffraction data sets were collected on SPring-8 beamlines. The best crystal of RVV-V generated data sets to 1.9,Å resolution. [source] | ||||||||||