High-resolution structure of proteinase K cocrystallized with digalacturonic acid

The crystal structure of proteinase K cocrystallized with digalacturonic acid and HEPES was solved at 1.32 Å resolution. The disaccharide was rotationally disordered about a twofold axis, but nonetheless contributed to intermolecular lattice interactions.

Structure of a D-tagatose 3-epimerase-related protein from the hyperthermophilic bacterium Thermotoga maritima

The crystal structure of a hyperthermophilic D-tagatose 3-epimerase-related protein with a unique active-site architecture was determined.

The structure of NMB1585, a MarR-family regulator from Neisseria meningitidis

The structure of the MarR-family regulator NMB1585 from N. meningitidis has been solved using data extending to 2.1 Å resolution.

Cloning, purification, crystallization and preliminary X-ray analysis of a chimeric NADPH-cytochrome P450 reductase

A 2.5 Å resolution data set was collected from a crystal of a soluble chimeric form of NADPH-cytochrome P450 reductase (CPR) produced using a fusion gene composed of the yeast FMN and the human FAD domains. The chimeric protein was crystallized in a modified conformation compared with the previously solved structures.

Crystallization and preliminary X-ray diffraction analysis of the lectin from Canavalia boliviana Piper seeds

Canavalia boliviana lectin (Cbol) was purified using a Sephadex G-50 column and crystallized in the presence of X-Man by hanging-drop vapour diffusion at 293 K. After optimization, crystals suitable for diffraction were obtained using 0.1 M HEPES pH 7.5 and 3.0 M sodium formate.

Purification, crystallization and preliminary X-ray analysis of an amino­acylhistidine dipeptidase (PepD) from Vibrio alginolyticus

The aminoacylhistidine dipeptidase encoded by V. alginolyticus pepD has been overexpressed and crystallized.

Crystallization and preliminary crystallographic analysis of thermophilic cellulase from Fervidobacterium nodosum Rt17-B1

The thermophilic cellulase FnCel5A from F. nodosum Rt17-B1 has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution for the native enzyme and to 1.7 Å resolution for the selenomethionyl derivative.

Crystallization and data collection of the nucleotide-binding domain of Mg-ATPase

Recombinant nucleotide-binding domain of E. coli Mg-ATPase was expressed, purified and used to produce crystals that diffracted to 1.53 Å resolution.

Expression, purification and crystallization of class C acid phosphatases from Francisella tularensis and Pasteurella multocida

Two members of the class C family of bacterial nonspecific acid phosphatases have been cloned, expressed, purified and crystallized. One of the crystal forms exhibited epitaxial twinning.

A preliminary neutron crystallographic study of an A-DNA crystal

The LADI-III diffractometer at the Institut Laue–Langevin has been used to carry out the first neutron crystallographic study of a DNA oligonucleotide in the A conformation. The crystal size was 0.06 mm³, the smallest ever used successfully for a study of this type. The results provide evidence of unexpected base protonation and illustrate the opportunities that now exist for nucleic acid crystallography using both hydrogenated and perdeuterated oligonucleotides.

Site-specific unglycosylation to improve crystallization of the metabotropic glutamate receptor 3 extracellular domain

The site-specific unglycosylated mutant of metabotropic glutamate receptor 3 extracellular domain has been overexpressed, purified, and crystallized. A complete data set has been collected to 2.35 Å resolution.

Crystallization and preliminary X-ray diffraction analyses of several forms of the CfaB major subunit of enterotoxigenic Escherichia coli CFA/I fimbriae

Three fusion proteins were generated in order to resolve the atomic structure of the CFA/I fimbriae of enterotoxigenic E. coli. CfaEB is a fusion of the minor and major CFA/I subunits, while CfaBB and CfaBBB are tandem fusions of two and three repeats, respectively, of the major subunit. Each protein was crystallized and the crystal structures of each of these fusions were determined successively by the molecular-replacement method using the CfaE crystal structure as an initial phasing model.

Crystallization and preliminary characterization of the Thermus thermophilus RNA helicase Hera C-terminal domain

Two C-terminal fragments of the RNA helicase Hera have been crytallized in three crystal forms, one of which was phased by MAD using a single selenium site.

Crystallization and preliminary X-ray analysis of di­hydrodipicolinate synthase from Clostridium botulinum in the presence of its substrate pyruvate

Dihydrodipicolinate synthase (DHDPS) catalyzes an important step in lysine biosynthesis. Here, the crystallization and preliminary diffraction analysis to 1.2 Å resolution of DHDPS from C. botulinum in the presence of its substrate pyruvate is reported.

Expression, purification and crystallization of the cofactor-independent monooxygenase SnoaB from the nogalamycin biosynthetic pathway

The cofactor-independent monooxygenase SnoaB from the nogalamycin-biosynthesis pathway and a designed selenomethionine-substituted double mutant were crystallized in space group P2₁2₁2. The native enzyme diffracted to a resolution of 2.4 Å.

Expression, purification and preliminary diffraction studies of CmlS

CmlS from S. venezuelae is a flavin-dependent halogenase that is involved in the biosynthesis of the widely used antibiotic chloramphenicol. Here, the crystallization of CmlS and analysis of the initial diffraction data are reported.

Crystallization and preliminary X-ray diffraction analysis of a putative two-domain-type laccase from a metagenome

A putative two-domain-type laccase from a metagenome was crystallized using the sitting-drop vapour-diffusion method. Diffraction data were collected to a resolution of 1.7 Å.

Crystallization and preliminary X-ray crystallographic studies of the Z-DNA-binding domain of a PKR-like kinase (PKZ) in complex with Z-DNA

The Z-DNA-binding domain of a PKR-like kinase (PKZ) from goldfish was crystallized in complex with d(TCGCGCG)₂. The crystals belonged to space group C2 and diffracted to 1.7 Å resolution.

Preliminary crystallographic study of two cuticle-degrading proteases from the nematophagous fungi Lecanicillium psalliotae and Paecilomyces lilacinus

Two cuticle-degrading proteases Ver112 and PL646 were purified from the nematophagous fungi L. psalliotae and P. lilacinus, respectively. The protease Ver112 and a complex between PL646 and a tetrapeptide inhibitor were crystallized. Diffraction data were collected to 1.65 and 2.2 Å resolution, respectively.

Crystallization and preliminary X-ray analysis of a cohesin-like module from AF2375 of the archaeon Archaeoglobus fulgidus

A cohesin-like module from the hyperthermophilic archaeon A. fulgidus was cloned, expressed, purified and crystallized. X-ray diffraction data were collected to 1.82 Å resolution.

Expression, purification, crystallization and preliminary crystallographic study of the carboxyl-terminal domain of the human voltage-gated proton channel Hv1

Here, the C-terminal domain of the human voltage-gated proton channel Hv1 (C-Hv1) was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method.

Preliminary X-ray crystallographic studies of Bacillus subtilis SpeA protein

In order to further illustrate the catalytic mechanism of arginine decarboxylase by determining the three-dimensional structure of the enzyme the speA gene was amplified from B. subtilis genomic DNA and cloned. The enzyme was expressed in Escherichia coli and purified to homogeneity by nickel-chelation chromatography followed by size-exclusion chromatography. High-quality crystals were obtained using the hanging-drop vapour-diffusion method at 298 K.

Cloning, expression, purification and preliminary crystallographic studies of the adenylate/uridylate-rich element-binding protein HuR complexed with its target RNA

An RNA-binding region of human HuR bound to an 11-base RNA fragment has been crystallized. The crystals diffracted to a resolution of 1.8 Å and belonged to space group P2₁2₁2₁.

Crystallization and preliminary X-ray analysis of human liver α-enolase

α-Enolase from human liver (hENO1) was expressed as a soluble protein and purified by affinity column chromatography and gel filtration. Crystals were obtained by the hanging-drop vapour-diffusion method and diffracted to 2.5 Å resolution.

Expression, purification and preliminary X-ray diffraction studies of VERNALIZATION1_208–341 from Arabidopsis thaliana

A C-terminal fragment of VERNALIZATION1 from A. thaliana, a DNA-binding protein required for the acceleration of flowering in response to prolonged cold treatment, was crystallized. X-ray diffraction data were collected to a resolution of 2.1 Å.

Crystallization and preliminary X-ray analysis of the tumour necrosis factor α–tumour necrosis factor receptor type 2 complex

The formation, crystallization and preliminary X-ray diffraction analysis of the tumour necrosis factor α (TNF)–tumour necrosis factor receptor type 2 (TNFR2) complex are described. The initial electron-density map, which was calculated using only the phases of refined TNF trimer structures, could detect the main chains and side chains of TNFR2 around the TNF trimer.

Purification, crystallization and preliminary X-ray diffraction of wild-type and mutant recombinant human transforming growth factor β-induced protein (TGFBIp)

Wild-type and mutant recombinant human transforming growth factor β-induced protein (TGFBIp) were cloned, purified and crystallized. Preliminary X-ray crystallography data were obtained from wild-type TGFBIp.

Expression, crystallization and preliminary crystallographic analysis of PilZ_XAC1133 from Xanthomonas axonopodis pv. citri

The cloning and expression of recombinant PilZ_XAC1133, a protein belonging to the PilZ superfamily, are described. PilZ proteins are associated with the control of several complex behaviours in bacteria and in some cases have been shown to bind c-diGMP. PilZ_XAC1133 containing selenomethionine produced crystals that diffracted to 1.85 Å resolution.

Crystallization and preliminary X-ray analysis of LipL32 from Leptospira interrogans serovar Copenhageni

Recombinant selenomethionine-labelled LipL32, the major surface protein of pathogenic Leptospira, has been purified and crystallized. Data sets from two crystals were collected, one of which diffracted to 2.25 Å resolution.

Purification, crystallization and preliminary crystallographic analysis of Est-Y29: a novel oligomeric β-lactamase

Est-Y29, a novel oligomeric class C β-lactamase from a metagenomic library, was crystallized in space group I4₁ and diffraction data were collected to 1.49 Å resolution.

Purification, crystallization and preliminary crystallographic study of low oxygen-affinity haemoglobin from cat (Felis silvestris catus) in two different crystal forms

Preliminary studies were carried out to purify and crystallize the sample from cat (Felis silvestris catus), a low oxygen-affinity haemoglobin in different crystal forms.

A preliminary neutron diffraction study of γ-chymotrypsin

A 2.0 Å resolution neutron diffraction data set has been collected from a D₂O-soaked γ-chymotrypsin crystal at low pH on the Institute Laue–Langevin LADI-III beamline.