scholarly journals Purification, crystallization and preliminary X-ray crystallographic studies of Rv3705c fromMycobacterium tuberculosis

2014 ◽  
Vol 70 (8) ◽  
pp. 1090-1092
Author(s):  
Feifei Lu ◽  
Feng Gao ◽  
Honglin Li ◽  
Weimin Gong ◽  
Lin Zhou ◽  
...  

The conserved protein Rv3705c fromMycobacterium tuberculosishas been cloned, expressed, purified and crystallized by the sitting-drop vapour-diffusion method using PEG 3350 as a precipitant. The Rv3705c crystals exhibited space groupP6122 orP6522, with unit-cell parametersa=b= 198.0,c= 364.1 Å, α = β = 90, γ = 120°, and diffracted to a resolution of 3.3 Å.

1999 ◽  
Vol 55 (4) ◽  
pp. 918-920 ◽  
Author(s):  
Gurvan Michel ◽  
Tristan Barbeyron ◽  
Didier Flament ◽  
Thierry Vernet ◽  
Bernard Kloareg ◽  
...  

A recombinant form of His-tagged κ-carrageenase from Pseudoalteromonas carrageenovora has been expressed, purified and crystallized. Crystals have been obtained by the vapour-diffusion method using polyethylene glycol (Mr = 4000) as a precipitant. These crystals belong to the space group P212121, with unit-cell parameters a  =  58.2, b  =  62.8, c  =  77.9 Å, and diffract to 2.2 Å resolution on a rotating-anode X-ray source.


Author(s):  
Fang Lu ◽  
Bei Zhang ◽  
Yong Liu ◽  
Ying Song ◽  
Gangxing Guo ◽  
...  

Phytases are phosphatases that hydrolyze phytates to less phosphorylatedmyo-inositol derivatives and inorganic phosphate. β-Propeller phytases, which are very diverse phytases with improved thermostability that are active at neutral and alkaline pH and have absolute substrate specificity, are ideal substitutes for other commercial phytases. PhyH-DI, a β-propeller phytase fromBacillussp. HJB17, was found to act synergistically with other single-domain phytases and can increase their efficiency in the hydrolysis of phytate. Crystals of native and selenomethionine-substituted PhyH-DI were obtained using the vapour-diffusion method in a condition consisting of 0.2 Msodium chloride, 0.1 MTris pH 8.5, 25%(w/v) PEG 3350 at 289 K. X-ray diffraction data were collected to 3.00 and 2.70 Å resolution, respectively, at 100 K. Native PhyH-DI crystals belonged to space groupC121, with unit-cell parametersa = 156.84,b = 45.54,c = 97.64 Å, α = 90.00, β = 125.86, γ = 90.00°. The asymmetric unit contained two molecules of PhyH-DI, with a corresponding Matthews coefficient of 2.17 Å3 Da−1and a solvent content of 43.26%. Crystals of selenomethionine-substituted PhyH-DI belonged to space groupC2221, with unit-cell parametersa = 94.71,b= 97.03,c= 69.16 Å, α = β = γ = 90.00°. The asymmetric unit contained one molecule of the protein, with a corresponding Matthews coefficient of 2.44 Å3 Da−1and a solvent content of 49.64%. Initial phases for PhyH-DI were obtained from SeMet SAD data sets. These data will be useful for further studies of the structure–function relationship of PhyH-DI.


Author(s):  
Hye Jin Kim ◽  
Eunha Hwang ◽  
Young-Hyun Han ◽  
Saehae Choi ◽  
Woo Cheol Lee ◽  
...  

NORE1 is an important tumour suppressor in human cancers that interacts with the pro-apoptotic protein kinase MST1/2 through SARAH domains. The SARAH domain (residues 366–413) of human NORE1 was expressed inEscherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystal diffracted to 2.7 Å resolution and belonged to space groupP6122, with unit-cell parametersa=b= 73.041,c = 66.092 Å, α = β = 90, γ = 120°.


2014 ◽  
Vol 70 (12) ◽  
pp. 1643-1645
Author(s):  
Stephanie S. Dawes ◽  
Sharon L. Kendall ◽  
Edward N. Baker ◽  
J. Shaun Lott

KstR2 (Rv3557c) is one of two TetR-family transcriptional repressors of cholesterol metabolism inMycobacterium tuberculosis. The ability to degrade cholesterol fully is important for pathogenesis, and therefore this repressor was expressed, purified and crystallized. Crystals of KstR2 diffracted to better than 1.9 Å resolution and belonged to space groupC2, with unit-cell parametersa = 72.3,b= 90.3,c= 49.7 Å, α = γ = 90, β = 128.2°.


1999 ◽  
Vol 55 (4) ◽  
pp. 904-906 ◽  
Author(s):  
Hanspeter Lauble ◽  
Klaas Decanniere ◽  
Harald Wajant ◽  
Siegfried Förster ◽  
Franz Effenberger

Hydroxynitrile lyase from M. esculenta (cassava) was crystallized in two different crystal forms by the hanging-drop vapour-diffusion method. Crystals of form I were obtained from a mixture of polyethylene glycol 8000 and 2-methyl-2,4-pentanediol, and belong to the tetragonal space group P41212 or its enantiomorph P43212, with unit-cell parameters a  =  b  =  105.9, c  =  188.9 Å and with two molecules in the asymmetric unit. These crystals diffract to 2.9 Å with conventional X-ray sources and beyond 2.1 Å resolution with synchrotron radiation. The crystals are relatively sensitive to radiation damage and conditions for flash-cooling the crystals have been established. A complete native data set has been collected up to 2.2 Å resolution. Crystal form II has been obtained at pH 5.6 using lithium sulfate as a precipitant. The crystals apparently belong to the orthorhombic space group P21212, with unit-cell parameters a = 117.52, b = 127.09 and c = 78.08 Å, have two molecules in the asymmetric unit and diffract to beyond 2.0 Å resolution. A complete native data set has been collected to 2.2 Å resolution.


1999 ◽  
Vol 55 (7) ◽  
pp. 1356-1358 ◽  
Author(s):  
Clare E. M. Williams ◽  
Daniel J. White ◽  
Laure Delarbre ◽  
Lesley A. Mitchenall ◽  
Richard N. Pau ◽  
...  

Crystals of the molbindin ModG (subunit Mr = 14359 Da), a cytoplasmic molybdate-binding protein from Azotobacter vinelandii, were grown by vapour diffusion. Both apo and tungstate-bound forms were crystallized and X-ray data were collected at 100 K. Apo-ModG crystallizes in space group P6322, with unit-cell dimensions a = b = 90.62, c = 79.46 Å. Native data to a resolution of 2.5 Å were collected from a single crystal, which showed a marked improvement in diffraction quality after annealing. Data from a single-site gold derivative were also collected at 2.7 Å resolution. Crystals of the ligand-bound form of ModG belong to space group P321, with unit-cell parameters a = b = 50.57, c = 79.29 Å. X-ray data to a resolution of 2.0 Å were collected.


1999 ◽  
Vol 55 (2) ◽  
pp. 544-546 ◽  
Author(s):  
M. Scott ◽  
R. W. Pickersgill ◽  
G. P. Hazlewood ◽  
H. J. Gilbert ◽  
G. W. Harris

Crystals of 1,5-α-arabinanase A from Pseudomonas fluorescens subspecies cellulosa have been obtained by vapour diffusion. The crystals belong to the space group P6122 with unit-cell parameters a = b = 91.6, c = 179.4 Å with one molecule in the asymmetric unit. The native crystals and, to a much greater extent, heavy-atom soaked crystals are sensitive to radiation which necessitates cryocooling. Suitable cryocooling conditions have been established, though a shrinkage of the unit cell is observed, with a = b = 88.8 and c = 176.9 Å.


Author(s):  
Yingjia Song ◽  
Jianghui Liu ◽  
De-Feng Li ◽  
Honglin Li ◽  
Shihua Wang ◽  
...  

Rv3899c, a hypothetical protein fromMycobacterium tuberculosisthat is conserved within the mycobacteria, is predicted to be secreted and has been found in culture filtrates. Here, Rv3899c has been cloned, expressed inEscherichia coliand purified using standard chromatographic techniques. The hanging-drop vapour-diffusion method with PEG 3350 as a precipitant was used to crystallize the protein. N-terminal sequencing results showed that the amino-acid sequence of the crystallized protein began with GATAG, indicating that it is a fragment containing residues 184–410 of Rv3899c. Rv3899c184–410crystals exhibited the symmetry of space groupP212121, with unit-cell parametersa= 49.88,b= 54.72,c= 75.52 Å, α = β = γ = 90°, and diffracted to a resolution of 1.90 Å.


Author(s):  
Natalia Pakharukova ◽  
Minna Tuittila ◽  
Sari Paavilainen ◽  
Anton Zavialov

The attachment of many Gram-negative pathogens to biotic and abiotic surfaces is mediated by fimbrial adhesins, which are assembledviathe classical, alternative and archaic chaperone–usher (CU) pathways. The archaic CU fimbrial adhesins have the widest phylogenetic distribution, yet very little is known about their structure and mechanism of assembly. To elucidate the biogenesis of archaic CU systems, structural analysis of the Csu fimbriae, which are used byAcinetobacter baumanniito form stable biofilms and cause nosocomial infection, was focused on. The major fimbriae subunit CsuA/B complexed with the CsuC chaperone was purified from the periplasm ofEscherichia colicells co-expressing CsuA/B and CsuC, and the complex was crystallized in PEG 3350 solution using the hanging-drop vapour-diffusion method. Selenomethionine-labelled CsuC–CsuA/B complex was purified and crystallized under the same conditions. The crystals diffracted to 2.40 Å resolution and belonged to the hexagonal space groupP6422, with unit-cell parametersa=b= 94.71,c = 187.05 Å, α = β = 90, γ = 120°. Initial phases were derived from a single anomalous diffraction (SAD) experiment using the selenomethionine derivative.


2021 ◽  
pp. 1-3
Author(s):  
J. Maixner ◽  
J. Ryšavý

X-ray powder diffraction data, unit-cell parameters, and space group for tetrazene nitrate monohydrate, C2H9N11O4, are reported [a = 5.205(1) Å, b = 13.932(3) Å, c = 14.196(4) Å, β = 97.826(3)°, unit-cell volume V = 1019.8(4) Å3, Z = 4, and space group P21/c]. All measured lines were indexed and are consistent with the P21/c space group. No detectable impurities were observed.


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