First look at RNA in L-configuration

The first structure of an L-RNA duplex is compared with its D-RNA analogue.

Structure of the dodecamer r(GAUCACUUCGGU) with four 5′-overhang nucleotides

The structure of the RNA dodecamer demonstrates the stability of the mismatched base pairing conformation of UUCG in RNA double-helical structures in the crystalline state.

Substrate recognition and selectivity of plant glycerol-3-phosphate acyltransferases (GPATs) from Cucurbita moscata and Spinacea oleracea

The 1.55 Å resolution three-dimensional structure of a squash glycerol-3-phosphate acyltransferase (GPAT) was determined using the multiwavelength anomalous dispersion method. This structure analysis and chimeric studies of plant GPATs will enable us to elucidate the mechanism involved at the atomic level in a plant's tolerance of chilling temperature.

The application of multivariate statistical techniques improves single-wavelength anomalous diffraction phasing

The complex multivariate normal distribution is applied in a maximum-likelihood formalism to the single-wavelength anomalous diffraction experiment, producing better results compared with currently used programs in the test cases shown.

On the routine use of soft X-rays in macromolecular crystallography. Part II. Data-collection wavelength and scaling models

The magnitude of the anomalous signal of a xenon complex of porcine pancreatic elastase has been investigated as a function of data-collection wavelength and scaling protocol.

Flash-cooling of macromolecular crystals in a capillary to overcome increased mosaicity

The usefulness of flash-cooling in a capillary (FCC) is described. The FCC method cools crystals more slowly than conventional cooling and may be applied when conventional cooling using a cryoloop fails.

Structure of the snake-venom toxin convulxin

The crystal structure of the toxin convulxin from the venom of the South American rattlesnake C. durissus terrificus has been solved by molecular replacement and refined at 2.7 Å resolution. Analysis of the structure provides insights into the interactions between convulxin and platelet receptors.

Refinement of the structure of human Rab5a GTPase domain at 1.05 Å resolution

The structure of human Rab5a GTPase domain was refined at 1.05 Å resolution using the program SHELXL.

Structures of d(Gm⁵CGm⁵CGCGC) and d(GCGCGm⁵CGm⁵C): effects of methylation on alternating DNA octamers

The crystal structures of two DNA octamers with different positions of double methylated cytosines reinforce the idea that methylation of cytosine stabilizes the A-type conformation for alternating DNA sequences beginning with a 5′-purine.

Structure of an acidic phospholipase A₂ from Indian saw-scaled viper (Echis carinatus) at 2.6 Å resolution reveals a novel intermolecular interaction

The crystal structure of an acidic phospholipase A₂ from Indian saw-scaled viper (E. carinatus) has been determined at 2.6 Å resolution. It reveals an unusual intermolecular interaction involving the β-wing of one molecule and the calcium-binding loop of a symmetry-related molecule.

Phasing via SAD/MAD data: the method of the joint probability distribution functions

A general formula for phasing reflections via SAD/MAD data is provided; a procedure is described and successfully applied to various practical cases.

Structure of thaumatin in a hexagonal space group: comparison of packing contacts in four crystal lattices

Thaumatin packing contacts in four crystal lattices are described.

Structure of d(GCGAAAGC) (hexagonal form): a base-intercalated duplex as a stable structure

The crystal structure of a DNA fragment d(GCGAAAGC) shows that two octamers related by a crystallographic twofold symmetry are associated to form a base-intercalated duplex, which is maintained by two Watson-Crick G·C base pairs and a subsequent sheared G·A pair at both ends, the central four A residues being stacked on each other and exposed suggesting a functional role.

Structure and implications for the thermal stability of phosphopantetheine adenylyltransferase from Thermus thermophilus

The crystal structure of phosphopantetheine adenylyltransferasefrom T. thermophilus HB8 complexed with Ppant at 1.50 Å resolution is reported.

Crystallization of the effector-binding domains of BenM and CatM, LysR-type transcriptional regulators from Acinetobacter sp. ADP1

The effector-binding domains of two LysR-type transcriptional regulators, BenM and CatM, were crystallized. Additionally, the effector-binding domain of BenM was crystallized in the presence of effectors.

Cloning, expression, purification, crystallization and preliminary diffraction analysis of the C-terminal catalytic domain of human poly(ADP-ribose) polymerase

Two fragments of the C-terminal catalytic domain of human poly(ADP-ribose) polymerase (catPARP), Met-catPARP and Gly Ser-catPARP, were purified and crystallized. The crystals show diffraction to at least 3.5 Å resolution.

Preliminary crystallographic analysis of the NAC domain of ANAC, a member of the plant-specific NAC transcription factor family

Three crystal forms of the NAC domain of the A. thaliana ANAC protein have been obtained. One of the crystal forms is suitable for structure determination, which is being pursued by multiple isomorphous replacement.

Crystallization and preliminary X-ray analysis of 5′-methylthioribose kinase from Bacillus subtilis and Arabidopsis thaliana

Recombinant B. subtilis and A. thaliana MTR kinases were expressed, purified and crystallized. A colorimetric enzyme-activity assay and preliminary X-ray analysis of the crystals are described.

Crystallization and preliminary crystallographic analysis of β-hydroxyacyl ACP dehydratase (FabZ) from Plasmodium falciparum

The primary dehydratase of the fatty-acid elongation cycle, FabZ (β-hydroxyacyl ACP dehydratase), from P. falciparum has been crystallized.

Purification, crystallization and preliminary X-ray diffraction data of L7Ae sRNP core protein from Pyrococcus abyssii

L7Ae sRNP core protein from P. abyssi was crystallized and data were collected to 2.9 Å resolution.

Crystallization and preliminary X-ray diffraction study of the class A β-lactamase SED-1 and its mutant SED-G238C from Citrobacter sedlakii

The crystallization and preliminary X-ray study of the CTX-M-type extended-spectrum β-lactamase SED-1 from C. sedlakii and its mutant SED-G238C are reported.

Crystallization and preliminary X-ray crystallographic analysis of the N-terminal domain of XpsE protein from Xanthomonas campestris, an essential component of the type II protein-secretion machinery

The 21 kDa N-terminal domain of XpsE protein from X. campestris has been crystallized in the tetragonal space group P4₁2₁2 (or P4₃2₁2), with unit-cell parameters a = b = 56.1, c = 102.7 Å. A complete native data set to 2.0 Å has been collected.

Crystallization and preliminary X-ray analysis of tryptophan synthase α-subunits from Escherichia coli

To explore the molecular origin of the conformational stabilization mechanism of αTS, the α-subunit protein has been overexpressed in E. coli and crystallized using the hanging-drop vapour-diffusion method at 298 K.

Expression, purification, crystallization and preliminary crystallographic analysis of the diarrhoea-causing and virulence-determining region of rotaviral nonstructural protein NSP4

A fragment of the rotaviral enterotoxin NSP4 containing the viral virulence-determining peptide preceded by the diarrhoea-inducing coiled coil region has been crystallized. X-ray diffraction data have been collected to a resolution of 2.2 Å.

Co-crystallization of Leptospira interrogans peptide de­formylase with a potent inhibitor and molecular-replacement schemes with eight subunits in an asymmetric unit

Co-crystallization of peptide deformylase from Leptospira interrogans with its natural inhibitor actinonin produced diffraction-quality crystals that belong to space group P2₁, with unit-cell parameters a = 87.5, b = 119.1, c = 95.8 Å, β = 111.6°.

Crystallization of the crenarchaeal SRP core

The conserved ribonucleoprotein core of the signal recognition particle (SRP) has been crystallized. Both crystal forms are highly twinned and an explanation for the possible tetartohedral twinning is presented.

Crystallization and preliminary X-ray diffraction studies of DNA decamer d(CCAGGCCTGG) complexed with cobalt(III)-pepleomycin

An antitumour pepleomycin:d(CCAGGCCTGG) complex has been crystallized. The crystals belong to space group P2₁2₁2₁ and diffraction data have been collected to 2.8 Å resolution.

Crystallization and preliminary X-ray diffraction studies of the quorum-sensing regulator TraM from Agrobacterium tumefaciens

The quorum-sensing regulator TraM from A. tumefaciens has been crystallized and crystals diffract to 1.67 Å using synchrotron radiation. Selenomethionine-substituted TraM has also been crystallized and a two-wavelength MAD data set has been collected to 2.0 Å resolution.

A unique dye-decolorizing peroxidase, DyP, from Thanatephorus cucumeris Dec 1: heterologous expression, crystallization and preliminary X-ray analysis

X-ray crystallography of a unique dye-decolorizing peroxidase is reported.

Crystallization of Arabidopsis thaliana aceto­hydroxyacid synthase in complex with the sulfonylurea herbicide chlorimuron ethyl

The crystallization and preliminary X-ray diffraction analysis of the catalytic subunit of A. thaliana AHAS in complex with the sulfonylurea herbicide chlorimuron ethyl are reported.

Crystallization and X-ray analysis of the N-terminal core domain of a tumour-associated human DEAD-box RNA helicase, rck/p54

Crystals of Nc-rck/p54 have been grown and diffract to beyond 2 Å resolution.

Crystallization and preliminary X-ray crystallographic investigations on several thermostable forms of a Bacillus subtilis lipase

To understand the structural basis of thermostability, three crystal forms of B. subtilis lipase have been obtained at low pH along with three crystal forms of thermostable mutants generated by directed evolution.

Crystallization and preliminary X-ray characterization of Trichoderma reesei hydrophobin HFBII

Two crystal forms and the preliminary X-ray data of the T. reesei hydrophobin HFBII are described.

Expression, purification, crystallization and preliminary crystallographic analysis of a putative GTP-binding protein, YsxC, from Bacillus subtilis

YsxC, a putative GTP-binding protein from B. subtilis, has been crystallized in the presence and absence of GDP in forms suitable for structure determination.

Crystallization and preliminary X-ray diffraction analysis of the catalytic domain of recombinant human phosphodiesterase 3B

Three different diffraction-quality crystals of the catalytic domain of PDE3B have been obtained by the vapor-diffusion method. Structure solution is under way.

Crystallization of a carbamatase catalytic antibody Fab fragment and its complex with a transition-state analogue

Fab fragments of a mouse monoclonal carbamatase catalytic antibody have been prepared and crystallized in complex with a transition-state analogue in a form suitable for structure determination.

Crystallization of the oligopeptide-binding protein AppA from Bacillus subtilis

AppA, a receptor protein for oligopeptide uptake by B. subtilis, has been crystallized and native diffraction data to 2.3 Å spacing have been collected using synchrotron radiation.

Expression, purification, crystallization and preliminary X-ray studies of geranylgeranyl diphosphate synthase from Thermus thermophilus HB8

Geranylgeranyl diphosphate synthase and its SeMet derivative have been expressed in E. coli and crystallized in space group P4₁ or P4₃. Diffraction data from both crystals have been collected using the SPring-8 synchrotron-radiation source.

Crystallization and preliminary X-ray diffraction analysis of levansucrase (LsdA) from Gluconacetobacter diazotrophicus SRT4

Crystals of the G. diazotrophicus levansucrase have been obtained and diffract to 2.5 Å spacing using a synchrotron X-ray source.

The purification, crystallization and preliminary structural characterization of PhzF, a key enzyme in the phenazine-biosynthesis pathway from Pseudomonas fluorescens 2-79

PhzF, an enzyme that catalyzes a key condensation reaction in the biosynthesis of phenazine-1-carboxylic acid in P. fluorescens, was cloned, overexpressed and crystallized. Data collection from native and seleno-L-methionine-labelled crystals is reported.

Purification, characterization and preliminary crystallographic studies of a cysteine protease from Pachyrrhizus erosus seeds

The proteins Spe31 and Spe32 were purified simultaneously from the seeds of P. erosus. Crystals were obtained from the protein mixture by the hanging-drop vapour-diffusion method and they diffracted to a resolution of 2.61 Å on an in-house X-ray source. Gel electrophoresis under non-denaturing conditions showed that the protein crystallized was Spe31.

Crystallization of the pectate lyase PelI from Erwinia chrysanthemi and SAD phasing of a gold derivative

Crystals of pectate lyase from E. chrysanthemi have been obtained.  Diffraction data have been collected from a native crystal and a gold derivative.

Purification, crystallization and preliminary X-ray diffraction studies on pyruvate phosphate dikinase from maize

Crystals of pyruvate phosphate dikinase from maize have been obtained by the vapour-diffusion method. The crystals diffract to 2.3 Å using synchrotron radiation and belong to space group C2, with unit-cell parameters a = 108.2, b = 100.2, c = 108.4 Å, β = 96.5°.

Rhombohedral crystals of Mycobacterium tuberculosis phosphopantetheine adenylyltransferase

The crystallization of recombinant phosphopantetheine adenylyltransferase (PPAT), the penultimate enzyme in coenzyme A (CoA) biosynthesis, from M. tuberculosis is reported.

Crystallization and preliminary X-ray diffraction analysis of pyranose 2-oxidase from the white-rot fungus Trametes multicolor

Crystals of T. multicolor P2Ox have been grown from monomethylether PEG 2000, sodium acetate, MgCl₂ and Ta₆Br₁₂. They belonged to space group P2₁, with unit-cell parameters a = 99.9, b = 101.7, c = 135.6 Å, β = 90.85°.

W3Y single mutant of rubredoxin from Pyrococcus furiosus: a preliminary time-of-flight neutron study

Rubredoxin from the hyperthermophilic archaeon P. furiosus maintains its native structure at high temperatures (373 K). In order to investigate the role of hydrogen bonding, hydration and chain dynamics in this thermostability, wavelength-resolved Laue neutron diffraction data have been collected from the W3Y single mutant (Trp3→Tyr3) on the spallation neutron protein crystallography station (PCS) at Los Alamos Neutron Science Center.

An improved protocol for rapid freezing of protein samples for long-term storage

This paper describes a method for long-term storage of proteins for crystallography by freezing samples in PCR tubes. The use of PCR tubes results in more rapid freezing than the more traditional technique of dropping protein directly into liquid nitrogen and can successfully preserve samples that are damaged by the drop-freezing technique.

Notes for authors 2004