scholarly journals Crystallization and preliminary X-ray diffraction analysis ofHypocrea jecorinaCel7A in two new crystal forms

2014 ◽  
Vol 70 (6) ◽  
pp. 773-776 ◽  
Author(s):  
Annette M. Bodenheimer ◽  
Matthew J. Cuneo ◽  
Paul D. Swartz ◽  
Junhong He ◽  
Hugh M. O'Neill ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Crystal Form ◽  
Unit Cell ◽  
Carbohydrate Binding ◽  
X Rays ◽  
Cellobiohydrolase I ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Group ◽  
Cell Parameters

Cel7A (previously known as cellobiohydrolase I) fromHypocrea jecorinawas crystallized in two crystalline forms, neither of which have been previously reported. Both forms co-crystallize under the same crystallization conditions. The first crystal form belonged to space groupC2, with unit-cell parametersa= 152.5,b= 44.9,c= 57.6 Å, β = 101.2°, and diffracted X-rays to 1.5 Å resolution. The second crystal form belonged to space groupP6322, with unit-cell parametersa=b≃ 155,c≃ 138 Å, and diffracted X-rays to 2.5 Å resolution. The crystals were obtained using full-length Cel7A, which consists of a large 434-residue N-terminal catalytic domain capable of cleaving cellulose, a 27-residue flexible linker and a small 36-residue C-terminal carbohydrate-binding module (CBM). However, a preliminary analysis of the electron-density maps suggests that the linker and CBM are disordered in both crystal forms. Complete refinement and structure analysis are currently in progress.

Crystallization and preliminary X-ray study of two crystal forms of Klebsiella oxytoca diol dehydratase–cyanocobalamin complex

1999 ◽  
Vol 55 (4) ◽  
pp. 907-909 ◽  
Author(s):  
Jun Masuda ◽  
Tetsuya Yamaguchi ◽  
Takamasa Tobimatsu ◽  
Tetsuo Toraya ◽  
Kyoko Suto ◽  
...  
Keyword(s):  
Klebsiella Oxytoca ◽  
Crystal Form ◽  
Unit Cell ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Group ◽  
X Ray ◽  
Cell Parameters ◽  
Diol Dehydratase ◽  
Replacement Method

Two crystal forms of Klebsiella oxytoca diol dehydratase complexed with cyanocobalamin have been obtained and preliminary crystallographic experiments have been performed. The crystals belong to two different space groups, depending on the crystallization conditions. One crystal (form I) belongs to space group P212121 with unit-cell parameters a = 76.2, b = 122.3, c = 209.6 Å, and diffracts to 2.2 Å resolution using an X-ray beam from a synchrotron radiation source. The other crystal (form II) belongs to space group P21 with unit-cell parameters a = 75.4, b = 132.7, c = 298.8 Å, β = 91.9°, and diffracts to 3.0 Å resolution. For the purpose of structure determination, a heavy-atom derivative search was carried out and some mercuric derivatives were found to be promising. Structure analysis by the multiple isomorphous replacement method is now under way.

scholarly journals Crystallization of two operator complexes from theVibrio choleraeHigBA2 toxin–antitoxin module

2015 ◽  
Vol 71 (2) ◽  
pp. 226-233 ◽  
Author(s):  
San Hadži ◽  
Abel Garcia-Pino ◽  
Kenn Gerdes ◽  
Jurij Lah ◽  
Remy Loris
Keyword(s):  
Vibrio Cholerae ◽  
Crystal Form ◽  
Unit Cell ◽  
Dna Duplexes ◽  
Dna Duplex ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Group ◽  
Cell Parameters ◽  
Complex Crystals

The HigA2 antitoxin and the HigBA2 toxin–antitoxin complex fromVibrio choleraewere crystallized in complex with their operator box. Screening of 22 different DNA duplexes led to two crystal forms of HigA2 complexes and one crystal form of a HigBA2 complex. Crystals of HigA2 in complex with a 17 bp DNA duplex belong to space groupP3221, with unit-cell parametersa=b= 94.0,c= 123.7 Å, and diffract to 2.3 Å resolution. The second form corresponding to HigA2 in complex with a 19 bp duplex belong to space groupP43212 and only diffract to 3.45 Å resolution. Crystals of the HigBA2 toxin–antitoxin were obtained in complex with a 31 bp duplex and belonged to space groupP41212, with unit-cell parametersa=b= 113.6,c= 121.1 Å. They diffract to 3.3 Å resolution.

scholarly journals Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of the catalytic domain of a hyperthermostable endo-1,4-β-D-mannanase fromThermotoga petrophilaRKU-1

2010 ◽  
Vol 66 (9) ◽  
pp. 1078-1081 ◽  
Author(s):  
Camila Ramos Santos ◽  
Fabio Marcio Squina ◽  
Andreia Meza Navarro ◽  
Roberto Ruller ◽  
Rolf Prade ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Unit Cell ◽  
Unit Cell Parameters ◽  
Structural Determinants ◽  
Space Group ◽  
X Ray ◽  
Cell Parameters ◽  
Functional Studies ◽  
Thermotoga Petrophila ◽  
Potential Applications

Endo-1,4-β-D-mannanases play key roles in seed germination and fruit ripening and have recently received much attention owing to their potential applications in the food, detergent and kraft pulp industries. In order to delineate their structural determinants for specificity and stability, X-ray crystallographic investigations combined with detailed functional studies are being performed. In this work, crystals of the catalytic domain of a hyperthermostable endo-1,4-β-D-mannanase fromThermotoga petrophilaRKU-1 were obtained from three different conditions, resulting in two crystalline forms. Crystals from conditions with phosphate or citrate salts as precipitant (CryP) belonged to space groupP212121, with unit-cell parametersa= 58.76,b= 87.99,c= 97.34 Å, while a crystal from a condition with ethanol as precipitant (CryE) belonged to space groupI212121, with unit-cell parametersa= 91.03,b= 89.97,c= 97.89 Å. CryP and CryE diffracted to resolutions of 1.40 and 1.45 Å, respectively.

Crystallization of the catalytic domain of Clostridium cellulolyticum CeIF cellulase in the presence of a newly synthesized cellulase inhibitor

1998 ◽  
Vol 54 (1) ◽  
pp. 114-118 ◽  
Author(s):  
Corinne Reverbel-Leroy ◽  
Goetz Parsiegla ◽  
Vincent Moreau ◽  
Michel Juy ◽  
Chantal Tardif ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Glycosyl Hydrolase ◽  
Unit Cell ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Clostridium Cellulolyticum ◽  
Vapour Diffusion

The catalytic domain of the CeIF processive endocellulase, a family 48 glycosyl hydrolase from Clostridium cellulolyticum has been crystallized in the presence of a newly synthesized inhibitor (methyl 4-S-β-cellobiosyl-4-thio-β-cellobioside), by vapour diffusion, using PEG as a precipitant. The protein crystallizes in the orthorhombic P212121 space group and diffracts to a resolution of 2.0 Å. The unit-cell parameters are a = 61.4, b = 84.5, c = 121.9 Å.

scholarly journals Crystallization of the Ets1–Runx1–CBFβ–DNA complex formed on the TCRα gene enhancer

2014 ◽  
Vol 70 (10) ◽  
pp. 1380-1384 ◽  
Author(s):  
Masaaki Shiina ◽  
Keisuke Hamada ◽  
Taiko Inoue-Bungo ◽  
Mariko Shimamura ◽  
Shiho Baba ◽  
...  
Keyword(s):  
Crystal Packing ◽  
Cell Receptor ◽  
Unit Cell ◽  
X Rays ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Intrinsically Disordered ◽  
Binding Domains ◽  
Gene Enhancer

Gene transcription is regulated in part through the assembly of multiple transcription factors (TFs) on gene enhancers. To enable examination of the mechanism underlying the formation of these complexes and their response to a phosphorylation signal, two kinds of higher-order TF–DNA assemblies were crystallized composed of an unmodified or phosphorylated Ets1 fragment, a Runx1(L94K) fragment and a CBFβ fragment on the T-cell receptor (TCR) α gene enhancer. Within these complexes, the Ets1 and Runx1 fragments contain intrinsically disordered regulatory regions as well as their DNA-binding domains. Crystals of the complex containing unmodified Ets1 belonged to space groupP212121, with unit-cell parametersa= 78.7,b= 102.1,c= 195.0 Å, and diffracted X-rays to a resolution of 2.35 Å, and those containing phosphorylated Ets1 belonged to the same space group, with unit-cell parametersa= 78.6,b= 101.7,c= 194.7 Å, and diffracted X-rays to a similar resolution. To facilitate crystallization, a Runx1 residue involved in a hydrophobic patch that was predicted to be engaged in crystal packing based on the previously reported structures of Runx1-containing crystals was mutated.

scholarly journals Two crystal forms of a helix-rich fatty acid- and retinol-binding protein, Na-FAR-1, from the parasitic nematodeNecator americanus

2012 ◽  
Vol 68 (7) ◽  
pp. 835-838 ◽  
Author(s):  
Mads Gabrielsen ◽  
M. Florencia Rey-Burusco ◽  
Kate Griffiths ◽  
Andrew J. Roe ◽  
Alan Cooper ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Structural Information ◽  
Binding Protein ◽  
Crystal Form ◽  
Blood Feeding ◽  
Unit Cell ◽  
Retinol Binding Protein ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters

Na-FAR-1 is an unusual α-helix-rich fatty acid- and retinol-binding protein fromNecator americanus, a blood-feeding intestinal parasitic nematode of humans. It belongs to the FAR protein family, which is unique to nematodes; no structural information is available to date for FAR proteins from parasites. Crystals were obtained with two different morphologies that corresponded to different space groups. Crystal form 1 exhibited space groupP432 (unit-cell parametersa = b = c = 120.80 Å, α = β = γ = 90°) and diffracted to 2.5 Å resolution, whereas crystal form 2 exhibited space groupF23 (unit-cell parametersa = b = c = 240.38 Å, α = β = γ = 90°) and diffracted to 3.2 Å resolution. Crystal form 2 showed signs of significant twinning.

scholarly journals The hyperthermophilic cystathionine γ-synthase from the aerobic crenarchaeonSulfolobus tokodaii: expression, purification, crystallization and structural insights

2017 ◽  
Vol 73 (3) ◽  
pp. 152-158 ◽  
Author(s):  
Dan Sato ◽  
Tomoo Shiba ◽  
Sae Mizuno ◽  
Ayaka Kawamura ◽  
Shoko Hanada ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Gel Filtration ◽  
Unit Cell ◽  
X Ray Diffraction ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Group ◽  
Cell Parameters ◽  
Transsulfuration Pathway ◽  
Ph Conditions

Cystathionine γ-synthase (CGS; EC 2.5.1.48), a pyridoxal 5′-phosphate (PLP)-dependent enzyme, catalyzes the formation of cystathionine from an L-homoserine derivative and L-cysteine in the first step of the transsulfuration pathway. Recombinant CGS from the thermoacidophilic archaeonSulfolobus tokodaii(StCGS) was overexpressed inEscherichia coliand purified to homogeneity by heat treatment followed by hydroxyapatite and gel-filtration column chromatography. The purified enzyme shows higher enzymatic activity at 353 K under basic pH conditions compared with that at 293 K. Crystallization trials yielded three crystal forms from different temperature and pH conditions. Form I crystals (space groupP21; unit-cell parametersa= 58.4,b= 149.3,c= 90.2 Å, β = 108.9°) were obtained at 293 K under acidic pH conditions using 2-methyl-2,4-pentanediol as a precipitant, whereas under basic pH conditions the enzyme crystallized in form II at 293 K (space groupC2221; unit-cell parametersa= 117.7,b= 117.8,c= 251.3 Å) and in form II′ at 313 K (space groupC2221; unit-cell parametersa= 107.5,b= 127.7,c= 251.1 Å) using polyethylene glycol 3350 as a precipitant. X-ray diffraction data were collected to 2.2, 2.9 and 2.7 Å resolution for forms I, II and II′, respectively. Structural analysis of these crystal forms shows that the orientation of the bound PLP in form II is significantly different from that in form II′, suggesting that the change in orientation of PLP with temperature plays a role in the thermophilic enzymatic activity of StCGS.

Purification, crystallization and preliminary X-ray analysis of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa

1999 ◽  
Vol 55 (9) ◽  
pp. 1591-1593 ◽  
Author(s):  
G. Sainz ◽  
J. Vicat ◽  
R. Kahn ◽  
C. Tricot ◽  
V. Stalon ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Point Group ◽  
Unit Cell ◽  
Group Symmetry ◽  
Ornithine Carbamoyltransferase ◽  
Unit Cell Parameters ◽  
Crystal Forms ◽  
Space Group ◽  
Cell Parameters ◽  
Hexagonal Crystals

The catabolic ornithine carbamoyltransferase (OTCase) from Pseudomonas aeruginosa exhibits allosteric behaviour, with two conformational states of the molecule: an active R form and an inactive T form. The enzyme is a dodecamer with a molecular mass of 455700 Da. Three crystal forms have been obtained. Crystals of allosteric state T are rhombohedral, belonging to the R3 space group, with hexagonal unit-cell parameters a = b = 180.6, c = 122.0 Å. They diffract to a resolution of 4.5 Å. Two crystal forms for allosteric state R have been obtained, with hexagonal and cubic symmetries. Hexagonal crystals, which diffract to a resolution of 3.4 Å, belong to the space group P63 with unit-cell parameters a = b = 140.8, c = 145.6 Å. The cubic crystals belong to space group I23, with unit-cell parameter a = 134.32 Å and diffract to a resolution better than 2.5 Å. In all crystal forms, the dodecamer exhibits a 23 point-group symmetry.

scholarly journals Expression and crystallographic studies of theArabidopsis thalianaGDP-D-mannose pyrophosphorylase VTC1

2016 ◽  
Vol 72 (10) ◽  
pp. 795-798 ◽  
Author(s):  
Shun Zhao ◽  
Lin Liu
Keyword(s):  
Arabidopsis Thaliana ◽  
Ascorbic Acid ◽  
Vitamin C ◽  
Biosynthetic Pathway ◽  
Unit Cell ◽  
X Rays ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Ligand Free

GDP-D-mannose pyrophosphorylase catalyzes the production of GDP-D-mannose, an intermediate product in the plant ascorbic acid (AsA) biosynthetic pathway. This enzyme is a key regulatory target in AsA biosynthesis and is encoded byVITAMIN C DEFECTIVE 1(VTC1) in theArabidopsis thalianagenome. Here, recombinant VTC1 was expressed, purified and crystallized. Diffraction data were obtained from VTC1 crystals grown in the absence and presence of substrate using X-rays. The ligand-free VTC1 crystal diffracted X-rays to 3.3 Å resolution and belonged to space groupR32, with unit-cell parametersa=b= 183.6,c= 368.5 Å, α = β = 90, γ = 120°; the crystal of VTC1 in the presence of substrate diffracted X-rays to 1.75 Å resolution and belonged to space groupP21, with unit-cell parametersa= 70.8,b= 83.9,c= 74.5 Å, α = γ = 90.0, β = 114.9°.

scholarly journals Expression, purification and crystallization of the phosphate-binding PstS protein fromPseudomonas aeruginosa

2014 ◽  
Vol 70 (7) ◽  
pp. 906-910 ◽  
Author(s):  
Avi Neznansky ◽  
Yarden Opatowsky
Keyword(s):  
Crystal Form ◽  
Phosphate Transport ◽  
Unit Cell ◽  
Asymmetric Unit ◽  
Unit Cell Parameters ◽  
Orthorhombic Space Group ◽  
Solvent Content ◽  
Space Group ◽  
Cell Parameters ◽  
Crystal Volume

Pseudomonas aeruginosa(PA) infections pose a serious threat to human health. PA is a leading cause of fatal lung infections in cystic fibrosis and immune-suppressed patients, of sepsis in burn victims and of nosocomial infections. An important element in PA virulence is its ability to establish biofilms that evade suppression by the host's immune system and antibiotics. PstS, a periplasmic subunit of the Pst phosphate-transport system of PA, plays a critical role in the establishment of biofilms. In some drug-resistant PA strains, PstS is secreted in large quantities from the bacteria, where it participates in the assembly of adhesion fibres that enhance bacterial virulence. In order to understand the dual function of PstS in biofilm formation and phosphate transport, the crystal structure of PA PstS was determined. Here, the overexpression inEscherichia coliand purification of PA PstS in the presence of phosphate are described. Two crystal forms were obtained using the vapour-diffusion method at 20°C and X-ray diffraction data were collected. The first crystal form belonged to the centred orthorhombic space groupC2221, with unit-cell parametersa= 67.5,b= 151.3,c= 108.9 Å. Assuming the presence of a dimer in the asymmetric unit gives a crystal volume per protein weight (VM) of 2.09 Å3 Da−1and a solvent content of 41%. The second crystal form belonged to the primitive orthorhombic space groupP212121, with unit-cell parametersa= 35.4,b= 148.3,c= 216.7 Å. Assuming the presence of a tetramer in the asymmetric unit gives a crystal volume per protein weight (VM) of 2.14 Å3 Da−1and a solvent content of 42.65%. A pseudo-translational symmetry is present in theP212121crystal form which is consistent with a filamentous arrangement of PstS in the crystal lattice.

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