Optimization of crystallization conditions for biological macromolecules

Strategies and procedures for the optimization of macromolecular crystal-growth conditions are surveyed. The objective of optimization is to grow crystals with the greatest degree of perfection and that yield the most accurate and precise X-ray diffraction data.

New crystal structures of adenylate kinase from Streptococcus pneumoniae D39 in two conformations

Adenylate kinases play a critical role in intercellular homeostasis by the interconversion of ATP and AMP to two ADP molecules. Crystal structures of adenylate kinase from Streptococcus pneumoniae D39 (SpAdK) have been determined in ligand-free and inhibitor-bound states at 1.96 and 1.65 Å resolution, respectively.

Structure of the methano­furan/methanopterin-biosynthetic enzyme MJ1099 from Methanocaldococcus jannaschii

The structure of the methanofuran/methanopterin-biosynthetic enzyme (MJ1099) from M. jannaschii was determined at 1.7 Å resolution using anomalous scattering methods.

The structure of rice weevil pectin methylesterase

This article reports the structure of pectin methylesterase from rice weevil (Sitophilus oryzae) which is the first structure of an animal pectin methylesterase.

Structure of type II de­hydroquinase from Pseudomonas aeruginosa

The recombinant production and structural analysis of catalytically active type II dehydroquinase from P. aeruginosa are presented.

Structure of an Escherichia coli Hfq:RNA complex at 0.97 Å resolution

The crystal structure of the RNA chaperone Hfq in complex with RNA is presented at 0.97 Å resolution.

Ovine β-lactoglobulin at atomic resolution

The 1.12 Å resolution crystal structure of a triclinic form of β-lactoglobulin without ligand from the domestic sheep (O. aries) is reported.

Production, crystallization and preliminary X-ray diffraction of the Gαs α-helical domain in complex with a nanobody

A crystallization screen was set up for GαsAHD in complex with the specifically selected nanobody CA9177. A high-resolution (1.59 Å) data set was collected for one of the crystallization conditions.

Purification, crystallization and preliminary X-ray diffraction analysis of NADP-dependent glutamate de­hydrogenase from Aspergillus niger

NADP-dependent glutamate dehydrogenase from A. niger has been crystallized and its preliminary X-ray diffraction analysis at 2.9 Å resolution is reported.

Cloning, expression, purification, crystallization and preliminary X-ray characterization of allantoinase from Bacillus licheniformis ATCC 14580

The cloning, expression, purification, crystallization and preliminary X-ray data collection of allantoinase from B. licheniformis (AllBali) are presented.

Overproduction, crystallization and preliminary X-ray crystallographic analysis of Escherichia coli tRNA N⁶-threonylcarbamoyladenosine dehydratase

TcdA, a recently characterized E. coli tRNA N⁶-threonylcarbamoyladenosine dehydratase, previously called CsdL or YgdL, was crystallized in space group P2₁. Diffraction data were collected to 2.70 Å resolution using synchrotron radiation.

Crystallization and preliminary X-ray crystallographic analysis of an artificial molten-globular-like triosephosphate isomerase protein of mixed phylogenetic origin

The protein ccTIM is an engineered variant of the triosephosphate isomerase (TIM) enzyme and has sequence features from both prokaryotic and eukaryotic members of the TIM protein family. Moreover, it is as active as a native protein despite sharing some biophysical characteristics with a molten globular variant. This inscrutable protein was crystallized using the microbatch method and a full X-ray diffraction data set was collected to 2.2 Å resolution using a synchrotron-radiation source.

Purification, crystallization, preliminary X-ray diffraction and molecular-replacement studies of great cormorant (Phalacrocorax carbo) haemoglobin

The great cormorant hemoglobin has been isolated, purified and crystallized and the three dimensional structure is solved using molecular replacement technique.

Crystallization and preliminary X-ray diffraction data of β-galactosidase from Aspergillus niger

β-Galactosidase from A. niger has been expressed in S. cerevisiae, purified and crystallized in its deglycosylated form. X-ray data have been collected to 1.8 Å resolution.

In situ proteolysis, crystallization and preliminary X-ray diffraction analysis of a V_HH that binds listeria internalin B

V_HH R303 binds the Listeria virulence factor InlB. Crystals of R303 were obtained following in situ proteolysis with trypsin. Trypsin removed the flexible tag region of the protein, creating a homogeneous protein preparation. The crystals diffracted to 1.3 Å resolution.

Expression, purification, crystallization and preliminary X-ray crystallographic analysis of the extracellular olfactomedin domain of gliomedin

Gliomedin has an extracellular domain which mediates interactions between the myelinating Schwann cell and the axonal surface, and plays a role in the development and maintenance of the nodes of Ranvier. Here, the purification and crystallization of this olfactomedin domain are reported.

Purification, crystallization and preliminary crystallographic investigation of FrnE, a di­sulfide oxidoreductase from Deinococcus radiodurans

drFrnE, a disulfide oxidoreductase from D. radiodurans, was crystallized and preliminary diffraction data were obtained. The crystal belonged to space group P2₁22₁ and diffracted to 1.8 Å resolution.

Purification, crystallization and preliminary X-ray crystallographic analysis of the flagellar accessory protein FlaH from the methanogenic archaeon Methanocaldococcus jannaschii

The flagellar accessory protein FlaH from M. jannaschii has been purified and crystallized. The crystal structure has been solved using the MAD method.

Use of a novel microtitration protocol to obtain diffraction-quality crystals of 4-hy­droxy-2-oxoglutarate aldolase from Bos taurus

A novel vapour-diffusion crystallization approach was utilized to produce crystals of the enzyme 4-hydroxy-2-oxoglutarate aldolase. This `microtitration' method incorporated ammonium sulfate titration, a miniaturization of the working volume (<10 µl) and a controlled rate of evaporation by utilizing a large reservoir, and yielded crystals with favourable diffraction parameters compared with crystals obtained using the conventional hanging-drop method.

Purification, crystallization and preliminary X-ray diffraction analysis of pyridoxal kinase from Plasmodium falciparum (PfPdxK)

The cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of PdxK from P. falciparum are described.

Crystallization and preliminary X-ray diffraction studies of an L-amino-acid oxidase from Lachesis muta venom

An L-amino-acid oxidase from L. muta venom was crystallized and diffraction data were collected to 3.0 Å resolution.

Crystallization and preliminary X-ray analysis of the hypothetical deaminase RPB_0146 from Rhodopseudomonas palustris HaA2

The hypothetical deaminase RPB_0146 from R. palustris HaA2 was expressed in E. coli and purified. Crystallization and preliminarily X-ray crystallographic analysis of the recombinant deaminase were performed.

Crystallization and preliminary X-ray diffraction analysis of the S-adenosylhomocysteine hydrolase (SAHH) from Thermotoga maritima

Thermophilic S-adenosylhomocysteine hydrolase (SAHH) from Thermotoga maritima (TmSAHH), which catalyzes the reversible conversion of S-adenosylhomocysteine into adenosine and homocysteine, was expressed in Escherichia coli and crystallized.

Cloning, expression, purification, crystallization and X-ray crystallographic analysis of PhaA from Ralstonia eutropha

The PhaA protein from R. eutropha, the first enzyme in the polyhydroxybutyrate biosynthetic pathway, was crystallized and X-ray crystallographic analysis was performed.

Crystallization, room-temperature X-ray diffraction and preliminary analysis of Kaposi's sarcoma herpesvirus LANA bound to DNA

Rational mutagenesis of the latency-associated nuclear antigen residues 1008–1146 from Kaposi's sarcoma herpesvirus has allowed its crystallization in complex with DNA. A complete data set has been collected at room temperature at a synchrotron-radiation source.