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The pentapeptide repeat protein AlbG, provides self-resistance to the nonribosomally encoded hybrid polyketide-peptide termed albicidin. Analysis of the AlbG three-dimensional structure and the sequences of other pentapeptide repeat proteins that confer resistance to topiosomerase poisons suggests they have a similar dimer interface which may be critical to their interaction with topoisomerases.
TEM-72 is a quadruple mutant of TEM-1 and shows extended-spectrum β-lactamase properties. The present structure shows the presence of a citrate anion bound to the TEM-72 active site and supports the use of polycarboxylates as a scaffold for the design of broad-spectrum inhibitors of serine β-lactamases.
Crystals of SimR, a TetR-like efflux pump repressor from S. antibioticus, were obtained and X-ray data were recorded to a resolution of 2.3 Å.
The crystallization of an ASCH domain-containing protein from Z. mobilis ZM4 and the collection of diffraction data to 2.1 Å resolution are reported.
Biliverdin reductase (BVR) from Synechocystis sp. PCC6803 and its selenomethionine derivative were overexpressed and purified. X-ray diffraction data from an SeMet BVR microcrystal were collected to 3.0 Å resolution on microfocus beamline BL32XU at SPring-8.
Preliminary crystallographic analysis of the CBS domain protein MJ1004 from Methanocaldococcus jannaschii.
The preparative purification and crystallization of Flt3 receptor–ligand complexes are described.
The first high-angle neutron fibre diffraction results from an amyloid system are described. These were obtained using H2O/D2O isototopic replacement, and emphasise the ability of neutron fibre diffraction, used in conjunction with deuterium labelling approaches, to study novel aspects of amyloid conformation and hydration.
The Src-homology 2 (SH2) domain of Csk-family protein tyrosine kinases acts as a conformational switch to regulate their catalytic activity, which in turn promotes the inhibition of their proto-oncogenic targets, the Src-family kinases. Here, the expression, purification, small-angle X-ray scattering and preliminary diffraction analysis of the SH2 domain of the Csk-homologous kinase is reported.
Crystals of a chitinase from C. vernus were monoclinic, belonging to space group C2 with unit-cell parameters a = 172.3, b = 37.1, c = 126.4 Å, β = 127°, and diffracted to 2.1 Å resolution.
The whole extracellular regions of nectin-1 (nectin-1-EC) and nectin-2 (nectin-2-EC) were expressed in E. coli as inclusion bodies, solubilized in 8 M urea and then refolded by rapid dilution. Refolded nectin-1-EC and nectin-2-EC were subsequently purified using three chromatographic steps and crystallized by the hanging-drop vapour-diffusion method.
This work describes the purification and preliminary crystallographic analysis of the CBS-pair regulatory domain of the human ancient domain protein 4 (ACDP4), also known as CNNM4.
A lactonase from the hyperthermophilic archaeon S. islandicus has been crystallized. In combination with biochemical and bioengineering studies, it is expected that the structure of this protein will provide insight into the natural function of the phosphotriesterase-like lactonase family.
This work demonstrates how the digestion of a surface-exposed loop of B. fragilis type III glutamine synthetase results in crystal-packing rearrangements and explains how these changes facilitated the first structure solution.
Serine/threonine protein kinase SnRK2.6/OST1 (OPEN STOMATA 1) from A. thaliana has been crystallized by the hanging-drop vapour-diffusion method and a native data set has been collected to 2.8 Å resolution.
The crystallization of the C. perfringens delta-toxin is reported.
Juvenile hormone-binding protein and PBMHP-12, two major 30 kDa proteins, have been isolated and purified from the haemolymph of B. mori. The proteins were crystallized and the crystals diffracted X-rays to 2.9 and 1.3 Å resolution, respectively.
The DNA-binding domain of R. capsulatus MopB has been purified and crystallized for X-ray structure analysis.
The regulatory domain of M. tuberculosis aspartokinase, the enzyme which catalyses the first reaction step in the biosynthesis of the amino acids lysine, methionine and threonine, has been cloned, expressed, purified and crystallized. Preliminary X-ray diffraction analysis of several crystals revealed the presence of five distinct crystal forms.
Crystals of S. aureus MazF belong to space group P212121, diffract to 2.1 Å resolution and contain two MazF dimers in the asymmetric unit.
The cloning, expression, purification, crystallization and preliminary crystallographic analysis of mannosyl-3-phosphoglycerate phosphatase (MpgP) from T. thermophilus HB27 are reported. The stability of MpgP in solution was studied by size-exclusion chromatography and differential scanning fluorimetry assays.
A mutant of the haloalkane dehalogenase DhaA (DhaA31) from R. rhodochrous NCIMB 13064 and its complex with 1,2,3-trichloropropane were crystallized and the crystals diffracted to high resolution.
Crystallographic data of organic solvent-tolerant lipase from Bacillus sp. strain 42 was collected at 2.0 Å with unit-cell parameters a =117.41, b = 80.85, c = 99.44 Å, β=96.40°. The protein–solvent interactions will be studied since lipase 42 was stable in water-miscible solvent.
The expression, purification and crystallization of the SARS coronavirus nsp16 RNA-cap AdoMet-dependent (nucleoside-2′O)-methyltransferase in complex with its activating factor nsp10 are reported.
4-Coumarate:CoA ligase 2 from A. thaliana has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to a resolution of 1.6 Å.
A truncated version of pleckstrin encompassing the amino-terminal PH domain and the central DEP domain has been crystallized and preliminary diffraction analysis has been conducted. The quaternary structure and phospholipid-binding properties of this pleckstrin truncation were analyzed.
Crystals of a geranyl pyrophosphate methyltransferase in the biosynthetic pathway of the off-flavor terpenoid alcohol, 2-methylisoborneol were obtained in the absence and presence of cofactor, cofactor analog and substrate.
The effect of temperature during crystallization setup was analyzed and was found to influence precipitate formation and the crystal shape of the inactive E294A mutant of A. salmonicida ssp. achromogenes protease 1.