scholarly journals Structure of the MarR family protein Rv0880 fromMycobacterium tuberculosis

2015 ◽  
Vol 71 (6) ◽  
pp. 741-745 ◽  
Author(s):  
Yun-Rong Gao ◽  
Na Feng ◽  
Tao Chen ◽  
De-Feng Li ◽  
Li-Jun Bi
Keyword(s):  
Amino Acids ◽  
Mycobacterium Tuberculosis ◽  
Isoelectric Point ◽  
Unit Cell Parameters ◽  
Dimerization Domain ◽  
Winged Helix ◽  
Cell Parameters ◽  
Transcription Regulator ◽  
Pfam Database ◽  
Family Protein

Rv0880 from the pathogenMycobacterium tuberculosisis classified as a MarR family protein in the Pfam database. It consists of 143 amino acids and has an isoelectric point of 10.9. Crystals of Rv0880 belonged to space groupP1, with unit-cell parametersa= 54.97,b= 69.60,c= 70.32 Å, α = 103.71, β = 111.06, γ = 105.83°. The structure of the MarR family transcription regulator Rv0880 was solved at a resolution of 2.0 Å with anRcrystandRfreeof 21.2 and 24.9%, respectively. The dimeric structure resembles that of other MarR proteins, with each subunit comprising a winged helix–turn–helix domain connected to an α-helical dimerization domain.

Crystallization and preliminary X-ray diffraction studies on the N-utilizing substance-B (NusB) from Mycobacterium tuberculosis

2000 ◽  
Vol 56 (1) ◽  
pp. 64-66 ◽  
Author(s):  
B. Gopal ◽  
R. A. Cox ◽  
M. J. Colston ◽  
G. G. Dodson ◽  
S. J. Smerdon ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Ribosomal Rna ◽  
Diffusion Method ◽  
X Ray Diffraction ◽  
Hanging Drop ◽  
Core Complex ◽  
Unit Cell Parameters ◽  
Data Set ◽  
X Ray ◽  
Cell Parameters

N-utilizing substance B (NusB) is a protein which forms part of a complex assembly in transcriptional antitermination in Mycobacterium tuberculosis. It forms a heterodimer with the product of the NusE gene (identical to the ribosomal protein S10) and mediates the process of transcriptional antitermination by forming the core complex with the nut site of the ribosomal RNA along with other protein factors. NusB has been cloned and overexpressed in Escherichia coli and crystallized using the hanging-drop vapour-diffusion method. The space group is P212121, with unit-cell parameters a = 46.6, b = 64.2, c = 90.1 Å. A native data set complete to 1.6 Å resolution has been collected from a single crystal.

scholarly journals Preliminary X-ray crystallographic studies of the TIR domain of human Toll-like receptor 6

2014 ◽  
Vol 70 (8) ◽  
pp. 1053-1055 ◽  
Author(s):  
Tae-ho Jang ◽  
Hyun Ho Park
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Immune Response ◽  
Innate Immune ◽  
Toll Like Receptor ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Tir Domain

Toll-like receptor (TLR) proteins have been identified and shown to play a role in the innate immune response. TLR6 associated with TLR2 can recognize diacylated lipoprotein. In this study, the human TLR6 TIR domain corresponding to amino acids 640–796 was overexpressed inEscherichia coliusing engineered C-terminal His tags. The TLR6 TIR domain was then purified to homogeneity and crystallized at 20°C. Finally, X-ray diffraction data were collected to a resolution of 2.2 Å from a crystal belonging to space groupC2, with unit-cell parametersa= 127.60,b= 44.20,c= 75.72 Å, β = 118.89°

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 ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Unit Cell ◽  
Diffusion Method ◽  
Unit Cell Parameters ◽  
Space Group ◽  
X Ray ◽  
Cell Parameters ◽  
Vapour Diffusion

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.

scholarly journals Crystallization and preliminary crystallographic analysis of defective pollen wall (DPW) protein fromOryza sativa

2014 ◽  
Vol 70 (6) ◽  
pp. 758-760 ◽  
Author(s):  
Wei Wang ◽  
Yuanyuan Ma ◽  
Yang Suo ◽  
Liming Yan ◽  
Dabing Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Oryza Sativa ◽  
Crystallographic Analysis ◽  
Pollen Wall ◽  
X Rays ◽  
Unit Cell Parameters ◽  
Gene Encoding ◽  
Cell Parameters ◽  
Alcohol Synthesis

Thedefective pollen wall(dpw) gene ofOryza sativaencodes a fatty acid reductase (DPW) which plays important roles in primary fatty alcohol synthesis. DPW catalyzes the synthesis of 1-hexadecanol. The enzyme shows a higher specificity for palmitoyl-ACP than for palmitoyl-CoA as the substrate, and can only use NADPH as the cofactor. To gain an understanding of the molecular mechanism underlying the reaction catalyzed by DPW, the gene encoding DPW without the N-terminal 80 amino acids (DPWΔ80) was cloned into pET-28a vector and was overexpressed inEscherichia coli. DPWΔ80 was purified to homogeneity and screened for crystallization. DPWΔ80 in complex with NADPH produced crystals that diffracted X-rays to a resolution of 3.4 Å. The crystals belonged to space groupP61orP65, with unit-cell parametersa=b= 222.8,c= 114.0 Å, α = β = 90, γ = 120°.

scholarly journals Crystallization of the C-terminal domain of the bacteriophage T7 fibre protein gp17

2012 ◽  
Vol 68 (2) ◽  
pp. 166-171 ◽  
Author(s):  
Carmela Garcia-Doval ◽  
Mark J. van Raaij
Keyword(s):  
Amino Acids ◽  
Protein A ◽  
Anomalous Dispersion ◽  
Unit Cell ◽  
Bacteriophage T7 ◽  
Asymmetric Unit ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Mercury Derivative

Bacteriophage T7 attaches to its host using the C-terminal domains of its six fibres, which are trimers of the gp17 protein. A C-terminal fragment of gp17 consisting of amino acids 371–553 has been expressed, purified and crystallized. Crystals of two forms were obtained, belonging to space groupP212121(unit-cell parametersa= 61.2,b= 86.0,c= 118.4 Å) and space groupC2221(unit-cell parametersa= 68.3,b= 145.6,c= 172.1 Å). They diffracted to 1.9 and 2.0 Å resolution, respectively. Both crystals are expected to contain one trimer in the asymmetric unit. Multiwavelength anomalous dispersion phasing with a mercury derivative is in progress.

scholarly journals Crystallization and X-ray diffraction analysis of chondroitin lyase from baculovirus: envelope protein ODV-E66

2012 ◽  
Vol 68 (2) ◽  
pp. 190-192 ◽  
Author(s):  
Yoshirou Kawaguchi ◽  
Nobuo Sugiura ◽  
Momo Onishi ◽  
Koji Kimata ◽  
Makoto Kimura ◽  
...  
Keyword(s):  
Amino Acids ◽  
Diffraction Analysis ◽  
Envelope Protein ◽  
Unique Characteristic ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
X Ray ◽  
Cell Parameters ◽  
Baculovirus Infection

Baculovirus envelope protein ODV-E66 (67–704), in which the N-terminal 66 amino acids are truncated, is a chondroitin lyase. It digests chondroitin and chondroitin 6-sulfate efficiently, but does not digest chondroitin 4-sulfate. This unique characteristic is useful for the preparation of specific chondroitin oligosaccharides and for investigation of the mechanism of baculovirus infection. ODV-E66 (67–704) was crystallized; the crystal diffracted to 1.8 Å resolution and belonged to space groupP62orP64, with unit-cell parametersa = b = 113.5,c= 101.5 Å. One molecule is assumed to be present per asymmetric unit, which gives a Matthews coefficient of 2.54 Å3 Da−1.

scholarly journals Purification, crystallization and preliminary X-ray crystallographic studies of KstR2 (ketosteroid regulatory protein) fromMycobacterium tuberculosis

2014 ◽  
Vol 70 (12) ◽  
pp. 1643-1645
Author(s):  
Stephanie S. Dawes ◽  
Sharon L. Kendall ◽  
Edward N. Baker ◽  
J. Shaun Lott
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Cholesterol Metabolism ◽  
Regulatory Protein ◽  
Unit Cell ◽  
Transcriptional Repressors ◽  
Unit Cell Parameters ◽  
Space Group ◽  
X Ray ◽  
Cell Parameters ◽  
Better Than

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°.

scholarly journals Expression, purification and preliminary crystallographic analysis ofMycobacterium tuberculosisCysQ, a phosphatase involved in sulfur metabolism

2014 ◽  
Vol 70 (6) ◽  
pp. 750-753 ◽  
Author(s):  
Anna I. Erickson ◽  
Reta D. Sarsam ◽  
Andrew J. Fisher
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Sulfur Metabolism ◽  
Crystallographic Analysis ◽  
Transfer Reactions ◽  
Asymmetric Unit ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
Orthorhombic Space Group ◽  
X Ray ◽  
Cell Parameters

CysQ is part of the sulfur-activation pathway that dephosphorylates 3′-phosphoadenosine 5′-monophosphate (PAP) to regenerate adenosine 5′-monophosphate (AMP) and free phosphate. PAP is the product of sulfate-transfer reactions from sulfotransferases that use the universal sulfate donor 3′-phosphoadenosine 5′-phosphosulfate (PAPS). In some organisms PAP is also the product of PAPS reductases that reduce sulfate from PAPS to sulfite. CysQ fromMycobacterium tuberculosis, which plays an important role in the biosynthesis of sulfated glycoconjugates, was successfully purified and crystallized in 24% PEG 1500, 20% glycerol. X-ray diffraction data were collected to 1.7 Å resolution using a synchrotron-radiation source. Crystals grew in the orthorhombic space groupP212121, with unit-cell parametersa= 40.3,b= 57.9,c= 101.7 Å and with one monomer per asymmetric unit.

Structure ofD-tyrosyl-tRNATyrdeacylase using home-source Cu Kα and moderate-quality iodide-SAD data: structural polymorphism and HEPES-bound enzyme states

2010 ◽  
Vol 66 (5) ◽  
pp. 584-592 ◽  
Author(s):  
Manickam Yogavel ◽  
Sameena Khan ◽  
Tarun Kumar Bhatt ◽  
Amit Sharma
Keyword(s):  
Amino Acids ◽  
Conformational Changes ◽  
Unit Cell ◽  
Structural Polymorphism ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Groups ◽  
Cell Parameters ◽  
Short Period ◽  
Sad Phasing

D-Tyrosyl-tRNATyrdeacylase (DTD) is an editing enzyme that removes D-amino acids from mischarged tRNAs. The crystal structure ofPlasmodium falciparumDTD (PfDTD) was determined using the iodide-SAD phasing method. Iodide-derivatized PfDTD crystals were obtained using the quick cryo-soaking procedure in which native crystals were soaked for a short period of 10–30 s in cryoprotectant solution containing 0.2–1 MNaI. Iodide-SAD data sets were collected to 3.3 and 2.74 Å resolution from PfDTD crystals that belonged to two different space groups,P43andP1, using an in-house X-ray copper-anode source. This is the first report to detail structure solution using low iodide anomalous signal, modest resolution and redundancy and average solvent content for SAD phasing of 984 and 1312 amino acids in the triclinicP1 and tetragonalP43space groups, respectively. A total of 85% and 56% of the residues were automatically built into the iodide-phased electron-density maps usingPHENIX AutoBuild. The structure of HEPES-bound PfDTD was subsequently determined by molecular replacement and refined to 2.83 Å resolution. The crystals obtained from various batches of crystallization trials of PfDTD exhibited polymorphism in terms of belonging to different crystal forms and space groups. Even within a given crystal system the unit-cell parameters showed high non-isomorphism. These packing variations were exploited in order to conduct a systematic study of conformational changes in PfDTD. It is shown that the disposition of a ten-residue insertion loop affects packing within the PfDTD crystals and seems to determine the non-isomorphism in unit-cell parameters. By tracking the changes in PfDTD unit cells, it was possible to map conformational differences within PfDTD that may be of significance for enzyme activity.

scholarly journals Cloning, purification, crystallization and preliminary X-ray diffraction analysis of mouse PACSIN 3 protein

2012 ◽  
Vol 68 (2) ◽  
pp. 159-162 ◽  
Author(s):  
Xiaoyun Bai ◽  
Geng Meng ◽  
Xiaofeng Zheng
Keyword(s):  
Protein Structures ◽  
Membrane Dynamics ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
Actin Reorganization ◽  
X Ray ◽  
Cell Parameters ◽  
Bar Domain ◽  
Cytoplasmic Proteins ◽  
Family Protein

PACSIN-family proteins are cytoplasmic proteins that have vesicle-transport, membrane-dynamics, actin-reorganization and microtubule activities. Here, the N-terminal F-BAR domain of mouse PACSIN 3, which contains 341 amino acids, was successfully cloned, purified and crystallized. The crystal of PACSIN 3 (1–341) diffracted to 2.6 Å resolution and belonged to space groupP21, with unit-cell parametersa= 46.9,b= 54.7,c= 193.7 Å, α = 90, β = 96.9, γ = 90°. These data should provide further information on PACSIN-family protein structures.

Sign in / Sign up

Export Citation Format

Share Document