Preliminary X-ray Characterization (preliminary + x-ray_characterization)

Distribution by Scientific Domains


Selected Abstracts


Crystallization and preliminary X-ray characterization of a novel calcium-binding protein AtCBL2 from Arabidopsis thaliana

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 6 2003
Masamichi Nagae
A new family of calcineurin B-like calcium-binding proteins has recently been identified in Arabidopsis thaliana. AtCBL2, a member of this family, has been crystallized in the presence of calcium ions using polyethylene glycol as a precipitant at 293,K. The crystals belong to space group C2221, with unit-cell parameters a = 83.9, b = 118.1, c = 49.1,┼. The asymmetric unit contains one molecule, with a VM of 2.36,┼3,Da,1 and a solvent content of 48%. Native diffraction data to 2.1,┼ resolution have been collected using synchrotron radiation at SPring-8. [source]


Isolation, purification and preliminary X-ray characterization of Cpn60-2 (65,kDa heat-shock protein) from Mycobacterium tuberculosis

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 9 2002
Noam Adir
Cpn60-2 is a member of a unique family of putative molecular chaperones homologous to GroEL (Cpn60) but of unknown function and found only in Mycobacterium tuberculosis and closely related species. Cpn60-2 has mainly been studied for its strong immunogenity. Here, the purification, crystallization and preliminary crystallographic analysis of M. tuberculosis Cpn60-2 are reported. The crystals belong to space group P2, with unit-cell parameters a = 57, b = 115.5, c = 81.5,┼, , = 95.5░, and contain a dimer in the asymmetric unit. The crystals diffract to 4.0,┼ using a Cu rotating-anode X-ray generator. [source]


Alliin lyase (alliinase) from garlic (Allium sativum): crystallization and preliminary X-ray characterization

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 8 2002
Linda J. W. Shimon
The enzyme alliinase has been isolated from garlic bulbs and crystallized. The crystals belong to space group P21, with unit-cell parameters a = 70.191, b = 127.006, c = 108.085,┼, , = 93.384░. They diffract to 2.2,┼ at liquid-nitrogen temperature. Analysis of the Patterson self-rotation function suggests that the crystals contain two dimeric molecules per asymmetric unit. [source]


Crystallization and preliminary X-ray characterization of a thermostable pectate lyase from Thermotoga maritima

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 4 2002
Michael A. McDonough
Pectate lyase is an enzyme involved in the degradation of the pectate portion of the primary plant cell wall. A recombinant pectate lyase from Thermotoga maritima where three of the four cysteine residues have been mutated (C132I, C156N, C194L) has been crystallized. Crystals of the same morphology and trigonal space group R3 with similar unit-cell parameters were obtained under two different conditions. The first, 0.3,M (NH4)H2PO4 pH 4.2, gave crystals with a maximum size of 0.4 Î 0.2 Î 0.2,mm in one week that diffracted to a resolution of 1.87,┼ and had unit-cell parameters a = b = 80.6, c = 148.8,┼. The second, 0.1,M sodium acetate, 6%(w/v) PEG 4000 pH 6.5, gave the same size crystals in two weeks that diffracted to a resolution of 2.1,┼ and had unit-cell parameters a = b = 80.0, c = 150.1,┼. [source]


Crystallization and preliminary X-ray analysis of the archaeosine tRNA-guanine transglycosylase from Pyrococcus horikoshii

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 11 2001
Ryuichiro Ishitani
The archaeosine tRNA-guanine transglycosylase from the hyperthermophilic archaeon Pyrococcus horikoshii was crystallized and preliminary X-ray characterization was performed. Single crystals were grown by the hanging-drop vapour-diffusion method, using sodium/potassium phosphate and sodium acetate as precipitants. The space group is P41212 or P43212, with unit-cell parameters a = b = 99.28,(14), c = 363.74,(56),┼. The cryocooled crystals diffracted X-rays beyond 2.2,┼ resolution using synchrotron şradiation from station BL44XU at SPring-8 (Harima). Selenoşmethionine-substituted protein crystals were prepared in order to solve the structure by the MAD phasing method. [source]


Crystallization of the cytotoxic domain of a ribosome-inactivating colicin in complex with its immunity protein

ACTA CRYSTALLOGRAPHICA SECTION D, Issue 12 2000
Stephen Carr
The complex between the ribonuclease domain of the ribosome-inactivating colicin E3 and its protein inhibitor, the cognate immunity Im3, has been crystallized and preliminary X-ray characterization has been performed. Single crystals of the 1:1 complex were grown from hanging-drop vapour-diffusion experiments using 2-propanol as a precipitant. The space group is P3121 or P3221, with unit-cell parameters a = b = 93.7, c = 76.2,┼. When cryocooled, these crystals diffract to a resolution of 2.4,┼. A search for suitable conventional heavy-atom derivatives was unsuccessful and so Im3 mutants containing engineered cysteine or methionine residues have been produced for mercury soaks and selenomethionine-labelling experiments, respectively. [source]


Crystallization and preliminary X-ray characterization of a catalytic and ATP-binding domain of a putative PhoR histidine kinase from the ,-radioresistant bacterium Deinococcus radiodurans

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 4 2010
S. Caria
The gene product of histidine kinase DR2244 (putative phoR) encoded by Deinococcus radiodurans has been suggested to be involved in the PhoR,PhoB two-component regulatory system. This two-component signalling system is activated upon phosphate starvation in several bacteria, including D. radiodurans. Single crystals were obtained from a recombinant preparation of the catalytic/ATP-binding (CA) domain of D. radiodurans PhoR (79,224) overexpressed in Escherichia coli. The crystals belonged to space group P212121, with unit-cell parameters a = 46.9, b = 81.8, c = 204.6,┼. The crystals contained six molecules in the asymmetric unit. Diffraction data were collected to 2.4,┼ resolution on beamline ID23-2 of the European Synchrotron Radiation Facility. [source]


Crystallization and preliminary X-ray characterization of the Skp1,Fbg3 complex

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 1 2010
Taichi Kumanomidou
F-box proteins are the substrate-recognition components of Skp1,Cullin1,F-box protein,Rbx1 (SCF) ubiquitin ligase complexes. Fbs1, an F-box protein, binds specifically to proteins modified with high-mannose oligosaccharides. Fbg3, another F-box protein, has 51% sequence identity to Fbs1. Although the residues that are necessary for binding to oligosaccharides are conserved between Fbs1 and Fbg3, Fbg3 does not bind glycoproteins. Skp1 and Fbg3 were co-expressed in Escherichia coli and their complex was purified to homogeneity and crystallized. Microseeding combined with the sandwiched hanging-drop technique improved the quality of the resulting crystals. The plate-shaped crystals belonged to space group P212121, with unit-cell parameters a = 34.1, b = 76.6, c = 193.9,┼ and one molecule per asymmetric unit. [source]


Crystallization and preliminary X-ray characterization of a PaaX-like protein from Sulfolobus solfataricus P2

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 8 2009
Yi Cao
PaaX is a global regulator of the phenylacetyl-coenzyme A catabolon that adjusts the expression of different operons to that of the paa -encoded central pathway. In this study, the PaaX-like protein from the hyperthermophilic archaeon Sulfolobus solfataricus P2 was successfully crystallized by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant. Diffraction data were obtained to a resolution of 3.0,┼ using synchrotron radiation at the Photon Factory. The crystal belonged to space group P321, with unit-cell parameters a = 86.4, b = 86.4, c = 105.5,┼. [source]


Crystallization and preliminary X-ray characterization of a glycerol dehydrogenase from the human pathogen Salmonella enterica serovar Typhimurium

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2009
A. T. Gonšalves
Glycerol dehydrogenase (GldA) encoded by the STM4108 gene (gldA) has been related to the synthesis of HilA, a major transcriptional regulator that is responsible for the expression of invasion genes in the human pathogen Salmonella enterica serovar Typhimurium. Single colourless crystals were obtained from a recombinant preparation of GldA overexpressed in Escherichia coli. They belonged to space group P2221, with unit-cell parameters a = 127.0, b = 160.1, c = 665.2,┼. The crystals contained a very large number of molecules in the asymmetric unit, probably 30,35. Diffraction data were collected to 3.5,┼ resolution using synchrotron radiation at the European Synchrotron Radiation Facility. [source]


Heterologous expression, crystallization and preliminary X-ray characterization of CcCel6C, a glycoside hydrolase family 6 enzyme from the basidiomycete Coprinopsis cinerea

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 2 2009
Yuma Kurakata
CcCel6C is a gene that encodes a glycoside hydrolase family 6 (GH6) enzyme in the Coprinopsis cinerea genome. In the evolutionary tree of GH6 enzymes, the encoded enzyme was closely related to Cel6B from Humicola insolens, previously called endoglucanase VI, while its amino-acid sequence revealed a region corresponding to the C-terminal active-site-enclosing loop typical of cellobiohydrolase II. Here, the crystallization of CcCel6C produced in Escherichia coli is reported. The square prismatic crystal belonged to the triclinic space group P1, with unit-cell parameters a = 44.04, b = 45.11, c = 48.90,┼, , = 77.81, , = 87.34, , = 68.79░. Diffraction data were collected to 1.6,┼ resolution. [source]


Expression, purification and preliminary X-ray characterization of dl -2-haloacid dehalogenase from Methylobacterium sp.

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2007

dl -2-Haloacid dehalogenase from Methylobacterium sp. CPA1 (dl -DEX Mb) is a unique enzyme that catalyzes the dehalogenation reaction without the formation of an ester intermediate. A recombinant form of dl -DEX Mb has been expressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystal belongs to the hexagonal space group P63, with unit-cell parameters a = b = 186.2, c = 114.4,┼. The crystals are likely to contain between four and eight monomers in the asymmetric unit, with a VM value of 4.20,2.10,┼3,Da,1. A self-rotation function revealed peaks on the , = 180░ section. X-ray data have been collected to 1.75,┼ resolution. [source]


Crystallization and preliminary X-ray characterization of 1,3-propanediol dehydrogenase from the human pathogen Klebsiella pneumoniae

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2007
D. Maršal
1,3-Propanediol dehydrogenase (1,3-PD-DH), encoded by the dhaT gene, is a key enzyme in the dissimilation process for converting glycerol to 1,3-propanediol in the human pathogen Klebsiella pneumoniae. Single colourless crystals were obtained from a recombinant preparation of 1,3-propanediol dehydrogenase overexpressed in Escherichia coli. The crystals belong to space group P21, with unit-cell parameters a = 91.9, b = 226.6, c = 232.6,┼, , = 92.9░. The crystals probably contain two decamers in the asymmetric unit, with a VM value of 3.07,┼3,Da,1 and an estimated solvent content of 59%. Diffraction data were collected to 2.7,┼ resolution using synchrotron radiation at the ID14-4 beamline of the European Synchrotron Radiation Facility. [source]


Crystallization and preliminary X-ray characterization of aminopeptidase N from Escherichia coli

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2006
Yuko Onohara
A recombinant form of aminopeptidase N (molecular weight 99,kDa) from Escherichia coli was crystallized by the hanging-drop vapour-diffusion method using ammonium sulfate as a precipitating agent. The crystals belong to the hexagonal space group P3121, with unit-cell parameters a = b = 120.5, c = 171.0,┼. The crystals are most likely to contain one molecule in the asymmetric unit, with a VM value of 3.62,┼3,Da,1. Diffraction data were collected to 2.0,┼ resolution using Cu,K, radiation from a rotating-anode X-ray generator. [source]