A modification of previously published protocols for the crystallization of homotetrameric R67 DHFR was used to crystallize a mixed wild-type and variant tandem dimer of R67 DHFR. Surprisingly, a fully wild-type crystal structure was obtained, apparently as a consequence of selective proteolytic degradation of the variant.
The crystal structure of the SH3 domain of the p85β subunit of human PI3K was determined to 2.0 Å resolution by molecular replacement. The overall structure is very similar to that of the p85α subunit of PI3K. The binding of two proline-rich ligand peptides to p85β SH3 was also characterized.
A high-resolution X-ray crystallographic study of the effects of solvent deuteration on the crystallization of proteinase K shows negligibly small degradations of the crystals owing to solvent deuteration and small structural differences between nondeuterated and deuterated crystals of proteinase K.
The crystal structure of the catalytic domain of ARF GTPase-activating protein (ARFGAP) of Plasmodium falciparum has been determined at 2.4 Å resolution and compared with the structures of mammalian ARFGAPs.
The assembly of bacterial outer membrane proteins is catalyzed by the BAM complex, which is made up of five proteins (BamA–E). Structural and bioinformatic analysis of the C-terminal domain of E. coli BamC (BamCC) suggests that a negatively charged conserved surface on BamCC may be an important protein–protein interaction site.
The expression, purification, crystallization and diffraction of two methyltransferases BT_2972 and BVU_3255 from two Bacteroides species of antibiotic-resistant pathogens from the human intestine are reported.
The extracellular part of the human epithelial cell-adhesion molecule (EpCAM; CD326) was cloned, mutated to prevent N-linked glycosylation, expressed in insect cells, purified and crystallized. Diffraction data were collected to 1.95 Å resolution.
SecDF is a multi-path membrane protein required for efficient protein translocation and integration via the Sec translocon. Crystals of the first periplasmic domain of SecDF, an essential element for SecDF function, diffracted X-rays to 2.3 Å resolution.
SiiE from S. enterica is a giant adhesion molecule with a total of 5559 amino acids that is specifically required for initial contact between the pathogen and polarized epithelial cells. Using limited proteolysis, a fragment of SiiE that encompasses Ig domains 50–52 was produced and crystallized and diffraction data were collected to 1.85 Å resolution.
Dihydrodipicolinate synthase (DHDPS) catalyzes the first committed step of the lysine-biosynthetic pathway in plants and bacteria. The cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis (to 2.5 Å resolution) of DHDPS2 from A. thaliana are reported.
A complex of the human DNA-repair proteins XRCC4 and XLF was crystallized under high-salt and extreme dehydration conditions to produce diffraction to 3.9 Å resolution. Initial phasing information was obtained from molecular replacement with single-wavelength anomalous diffraction using tantalum bromide clusters.
The macromolecular complex of ICP (inhibitor of cysteine proteases) from P. berghei and falcipain-2 from P. falciparum has been prepared and crystallized, and a diffraction data set has been collected to a resolution of 2.6 Å.
The cloning, purification, crystallization and preliminary X-ray diffraction analysis of a novel staphylococcal phage dUTPase is reported. This protein contains a specific polypeptide insertion that is potentially responsible for modulation of expression of superantigenicity island genes.
The PYD domain of human NALP3 was crystallized. The crystals were found to belong to the primitive monoclinic space group P21, with unit-cell parameters a = 42.03, b = 60.14, c = 51.61 Å, β = 107.40°. The crystals were obtained at 293 K and diffracted to a resolution of 1.7 Å.
Human transthyretin, a hormone-binding protein of high abundance in blood and cerebrospinal fluid, is intrinsically amyloidogenic. Preliminary results from a neutron diffraction analysis of this protein that may shed light on the mechanism of tetramer dissociation and successive fatal aggregation are presented.
Two orthologous putative tRNA methyltransferases from P. furiosus and T. thermophilus have been expressed, purified and crystallized. X-ray diffraction data were collected to 2.2 and 2.05 Å, respectively.
DndE is one of five essential proteins that are required for the DNA phosphorothioation process. However, its exact biochemical role in sulfur modification of DNA remains unclear. In this study, the DndE protein homologue from Salmonella enterica serovar Cerro 87 was overexpressed, purified and crystallized.
Four different L27PATJ–(L27N,L27C)Pals1–L27MALS constructs have been cloned, expressed, purified and crystallized. Crystals of tripartite complex 1 of L27PATJ–(L27N,L27C)Pals1–L27MALS diffracted to 2.05 Å resolution.
In this study, the hyperthermostable arginine-binding protein isolated from T. maritima has been crystallized in ligand-free and arginine-bound forms. Crystals of the apo and holo forms diffracted to 2.2 and 2.7 Å resolution, respectively.