Atomic resolution structure of Escherichia coli dUTPase determined ab initio

Derivatization with a mercury compound increases the diffraction limit of E. coli dUTPase crystals to atomic resolution. Data collection to 1.05 Å allows the structure to be solved from the position of the single Hg atom in the asymmetric unit. The final refined dUTPase derivative structure is described and compared with the native structure.

Avian haemoglobins and structural basis of high affinity for oxygen: structure of bar-headed goose aquomet haemoglobin

The structure of bar-headed goose aquomet haemoglobin showed quaternary structural differences from lowland animal haemoglobins; critical residue differences are located in the α₁β₁ interface.

Structure of C-phycocyanin from Spirulina platensis at 2.2 Å resolution: a novel monoclinic crystal form for phycobiliproteins in phycobilisomes

The monoclinic crystal structure of C-phycocyanin from S. platensis has been determined at 2.2 Å resolution. The unique side-by-side packing of two hexamers in the asymmetric unit makes it possible that lateral energy transfer takes place by the β155–β155 route.

Structure of the N-terminal domain of Yersinia pestis YopH at 2.0 Å resolution

The crystal structure of the N-terminal regulatory domain of the Y. pestis effector protein YopH has been determined at a resolution of 2.0 Å.

Solving a 300 kDa multimeric protein by low-resolution MAD phasing and averaging/phase extension

The structure of a homohexameric protein particle was solved by employing a tantalum bromide derivative of the crystals obtained. A low-resolution MAD experiment (three wavelengths) rendered phases that could be improved by averaging and extended by applying density modifications.

Stabilization of active-site loops in NH₃-dependent NAD⁺ synthetase from Bacillus subtilis

The order/disorder changes of active-site loops upon substrate binding are described and implications for substrate recognition and stabilization are shown, along with the effect of a higher crystallization pH.

Checking nucleic acid crystal structures

The program SFCHECK, which analyses the quality of the structure-factor data, and the agreement of those data with the atomic coordinates, is used to survey 105 nucleic acid crystal structures from the NDB.

X-ray diffraction and atomic force microscopy analysis of twinned crystals: rhombohedral canavalin

Hemihedrally twinned rhombohedral crystals of the plant seed protein canavalin were successfully refined. Their investigation by atomic force microscopy revealed an extensive defect structure that is likely to be a consequence of the twinning.

Laboratory multiple-crystal X-ray topography and reciprocal-space mapping of protein crystals: influence of impurities on crystal perfection

Laboratory double-axis multiple-crystal X-ray topography and triple-axis high angular resolution reciprocal-space mapping have been combined to investigate the influence of acetylated lysozyme upon lysozyme crystal perfection.

Crystallization and preliminary crystallographic data of fructose-1,6-bis­phosphatase from human muscle

Crystallization of human muscle fructose-1,6-bisphosphatase has yielded complete data to 2.04 Å.

Crystallization and preliminary X-ray analyses of quaternary, ternary and binary protein–DNA complexes with involvement of AML1/Runx-1/CBFα Runt domain, CBFβ and the C/EBPβ bZip region

AML1/Runx-1/CBFα(Runt)–CBFβ–C/EBPβ(bZip)–DNA, AML1/Runx-1/CBFα(Runt)–C/EBPβ(bZip)–DNA and AML1/Runx-1/CBFα(Runt)–DNA, three types of protein–DNA complexes, have been crystallized. The former two crystals diffracted to 3 Å resolution and the latter crystal diffracted to 2.65 Å resolution.

Crystallization and preliminary X-ray analysis of the C/EBPβ C-terminal region in complex with DNA

Two kinds of C-terminal fragments of human C/EBPβ (residues 259–345 and 259–336) were crystallized in complex with a 16 bp DNA fragment from the tom-1 promoter. These two types of crystals diffracted to 2.1 and 1.8 Å resolution, respectively.

Crystallization and preliminary X-ray studies of meso-2,3-butanediol de­hydrogenase from Klebsiella pneumoniae IAM1063

Two different crystal forms of meso-2,3-butanediol dehydrogenase from K. pneumoniae IAM1063 have been obtained. Crystal forms I and II belong to space groups C2 and P2₁ and diffract to 2.0 and 1.7 Å resolution, respectively.

Crystallization and preliminary X-ray study of haem-binding protein from the bloodsucking insect Rhodnius prolixus

Rhodnius haem-binding protein (RHBP) from the bloodsucking insect R. prolixus, a 15 kDa protein, has been crystallized using polyethylene glycol as a precipitant. The crystals belong to the space group P4₁₍₃₎2₁2 and diffract beyond 2.6 Å resolution.

Two different crystal forms of sorcin, a penta-EF-hand Ca²⁺-binding protein

Sorcin, a penta EF-hand Ca²⁺-binding protein, and its Ca²⁺-binding domain have been crystallized. The crystals of the entire molecule diffract poorly, but two crystal forms grown from the domain that lacks the first 32 amino acids diffract to 2.1 and 2.7 Å resolution.

Crystallization and preliminary crystallographic studies of a ribosomal protein L30e from the hyperthermophilic archaeon Thermococcus celer

Crystals of ribosomal protein L30e from the hyperthermophilic archaeon T. celer were obtained and data diffracting to 1.96 Å were collected using a Cu Kα X-ray source. The crystals belong to space group P6₁/P6₅, with unit-cell parameters a = b = 48.32, c = 86.42 Å.

Optimization of Met8p crystals through protein-storage buffer manipulation

Met8p was successfully crystallized by combining crystallization screening results with incremental adjustments to the protein storage buffer.

Crystallization of two gluta­thione S-transferases from an unusual gene family

Two glutathione S-transferase isozymes from the mosquito Anopheles dirus (AdGST1-3 and AdGST1-4) from an alternately spliced gene family have been expressed, purified and crystallized. Determination of these structures will advance our understanding of how these enzymes inactivate pesticides and the structural consequences of alternate splicing.

Crystallization and preliminary investigations on a telomeric repeat sequence C₄A₂C₄A₂

The space group and the unit-cell parameters of crystals of C₂A₂C₄A₂ and Patterson and molecular-replacement studies on the crystals suggest that the molecules assume a tetraplex structure.

Crystallization and preliminary crystallographic analysis of deoxyuridine 5′-triphosphate nucleotido­hydrolase from Bacillus subtilis

Four different crystal forms of B. subtilis dUTPase in unliganded and potentially liganded (with a substrate analogue) forms have been analysed using synchrotron radiation.

Crystallization and preliminary X-ray diffraction analysis of NADP-dependent glyceraldehyde-3-phosphate de­hydrogenase of Synechococcus PCC 7942

NADP-dependent glyceraldehyde-3-phosphate dehydrogenase of Synechococcus PCC 7942 was crystallized in two different forms using ammonium sulfate as a precipitant. Form I crystals belong to the hexagonal space group P6₅ and form II crystals belong to the monoclinic space group C2.

Preliminary X-ray crystallographic analysis of a Ca²⁺-binding protein human S100A1

S100A1, a Ca²⁺-binding protein from the S100 protein family, has been crystallized. The crystals were stable during exposure to X-rays and diffracted to 2.6 Å resolution in-house.

Crystallization and preliminary X-ray analysis of AlgS, a bacterial ATP-binding cassette (ABC) protein specific to macromolecule import

Hexahistidine-tagged AlgS of Sphingomonas sp. A1 was overexpressed in E. coli and crystallized. X-ray diffraction data to 3.2 Å have been collected from the native crystal.

Crystallization and quaternary structure analysis of an Lrp-like regulatory protein from the hyperthermophile Pyrococcus furiosus

The Lrp-like regulatory protein, LrpA, from P. furiosus has been crystallized in a form suitable for X-ray diffraction studies. An octameric assembly for the protein is suggested by its behaviour on gel-filtration columns and analysis of the self-rotation function.

Crystallization and preliminary crystallographic studies of RhoGDI in complex with the radixin FERM domain

Crystals of RhoGDI complexed with the FERM domain of radixin were obtained and belong to the space group P2₁2₁2, with unit-cell parameters a = 130.9, b = 151.2, c = 71.2 Å. Diffraction data to a resolution of 2.9 Å have been obtained using synchrotron radiation.

Crystallographic characterization of the radixin FERM domain bound to the cytoplasmic tail of the adhesion protein ICAM-2

Crystals of the complex between the FERM domain of radixin and the cytoplasmic tail of ICAM-2 were obtained. The binary complex crystals were found to belong to the space group P3₁21 or P3₂21, with unit-cell parameters a = b = 100.44, c = 99.49 Å, and diffract to 2.6 Å resolution.

Purification, crystallization and preliminary X-ray diffraction analysis of the yeast Sec12Δp protein, a guanine nucleotide-exchange factor involved in vesicle transport

A 38.5 kDa cytoplasmic domain of Sec12p from S. cerevisiae, a guanine nucleotide-exchange factor of a GTP-binding protein Sar1p, a key factor in vesicle transport from the endoplasmic reticulum to the Golgi apparatus, has been expressed in S. cerevisiae and in E. coli. The proteins have been purified to homogeneity and X-ray diffraction data sets have been collected from two crystal forms to 2.6 and 2.0 Å resolution.

The MinD protein from the hyperthermophilic archaeon Pyrococcus horikoshii: crystallization and preliminary X-ray analysis

The cell-division inhibitor MinD from P. horikoshii was crystallized and preliminary X-ray diffraction studies were performed.

Crystallization and preliminary X-ray analysis of the complex between human CTLA-4 and B7-2

The receptor-binding domain of human B7-2 and the disulfide-linked homodimer of human CTLA-4 have been produced in E. coli, purified and cocrystallized in a monoclinic form (space group P2₁). A single crystal was obtained that diffracted to 3.2 Å resolution.

Crystallization of a recombinant form of the complete sequence of human γ-interferon: characterization by small-angle X-ray scattering, mass spectrometry and preliminary X-ray diffraction studies

The crystallization conditions of a recombinant form of the complete sequence of human γ-interferon have been determined after studying the behaviour of this protein in solution by small-angle X-ray scattering (SAXS) as a function of pH and salt type.

Crystallization and preliminary crystallographic characterization of catechol-O-methyltransferase in complex with its cosubstrate and an inhibitor

Recombinant catechol-O-methyltransferase has been crystallized with a new tight-binding inhibitor. This enzyme is an important target in the treatment of Parkinson's disease.

Cloning, expression, purification and preliminary X-ray crystallographic studies of Escherichia coli Hsp100 ClpB nucleotide-binding domain 1 (NBD1)

The E. coli Hsp100 ClpB nucleotide-binding domain 1 (NBD1) has been cloned, purified and crystallized. The crystals diffracted to 1.8 Å.

Crystallization and preliminary X-ray studies of snake gourd lectin: homology with type II ribosome-inactivating proteins

Snake gourd seed lectin has been crystallized. Preliminary X-ray studies indicate the lectin to be homologous to type II ribosome-inactivating proteins.

New crystal forms of Trypanosoma cruzi dUTPase

Crystals of T. cruzi dUTPase in two crystals forms, both belonging to the hexagonal space group P6₃22, have been obtained. Form I is similar to crystals reported previously, but the crystals are of better quality and produced under different conditions, while form II has not been reported before.

Crystallization and preliminary X-ray diffraction studies of glycerol 3-phosphate cytidylyltransferase from Staphylococcus aureus

Glycerol 3-phosphate cytidylyltransferase from S. aureus has been cloned, expressed, purified and subsequently crystallized. Determination of crystallization conditions required a combination of three optimization strategies.

Subunit dissociation and reassociation leads to preferential crystallization of haemoglobin Bart's (γ₄) from solutions of human embryonic haemoglobin Portland (ζ₂γ₂) at low pH

Low-pH incubation of human embryonic haemoglobin Portland (ζ₂γ₂) has resulted in the preferential crystallization of haemoglobin Bart's (γ₄), as the result of dissociation of the embryonic heterotetramer and preferential formation of the homotetramer.

Amphiphiles modify the properties of detergent solutions used in crystallization of membrane proteins

The effect of the amphiphile heptanetriol on the critical micelle concentration and the cloud-point temperature was measured for several detergents commonly employed in the crystallization of membrane proteins.