issue contents
March 2013 issue
Cover illustration: Structure of Vibrio cholerae ribosome hibernation promoting factor (De Bari & Berry, p. 228).
structural communications
The crystal structures of two mutants at position 29 of the dimeric hemoglobin from Vitreoscilla are reported, together with a discussion of the significance of these mutations.
The high-resolution crystal structure of human Dim2/TXNL4B indicated that a region previously reported to be responsible for its interaction with Prp6 is unlikely to facilitate this interaction.
PDB reference: TXNL4B, 4in0
The X-ray crystal structure of ribosome hibernation promoting factor from V. cholerae has been determined at 2.0 Å resolution. The crystal was phased by two-wavelength MAD using cocrystallized cobalt.
PDB reference: ribosome hibernation promoting factor, 4hei
The structures of wild-type MntC and of an R116A mutant have been determined to 2.7 and 3.5 Å resolution, respectively. The role of Arg116 in the outer shell of the active site is described.
Open access
The crystal structure of inosine 5′-monophosphate dehydrogenase from P. aeruginosa has been determined to 2.25 Å resolution.
PDB reference: inosine 5′-monophosphate dehydrogenase, 3zfh
crystallization communications
In various muscles, the two heads of myosin together with part of its α-helical coiled coil make up the minimal unit that is regulated. Reported here, for the first time, is the growth of three-dimensional crystals of such a heavy meromyosin, purified from squid, and their X-ray diffraction to near-atomic resolution (up to ∼5 Å).
In this study, the L,D-transpeptidase LdtMt1 was cloned, expressed and crystallized. Multiwavelength anomalous dispersion experiments have been carried out to obtain experimental phases using data at 2.9 Å resolution from a selenomethionine derivative.
The soluble NTPDase1 from L. pneumophila was crystallized in six crystal forms and the structure was solved using a sulfur SAD approach.
Purification, crystallization and preliminary crystallographic analysis of the marine α-amylase AmyP
A new marine derived α-amylase AmyP, classified into a new subfamily of GH13, was recombinantly expressed, crystallized and preliminarily analyzed.
Crystallization and preliminary X-ray crystallographic studies of DnaJ from Streptococcus pneumoniae
DnaJ from Streptococcus pneumoniae (SpDnaJ) is involved in the infectious disease process and is being developed as a potential vaccine to prevent bacterial infection. Here the expression, purification, crystallization and preliminary crystallographic analysis of SpDnaJ are reported.
Two nanobodies generated against the VAR2CSA DBL6∊-FCR3 domain involved in pregnancy-associated malaria were selected, expressed, purified and crystallized.
The small terminase subunit from a lactococcal 936 bacteriophage (strain 454) has been expressed, purified, crystallized and X-ray diffraction data collected to 2.4 Å resolution.
Substrate-free cap-binding domain of influenza A virus H1N1 polymerase subunit PB2 has been crystallized to show the structural details and clarify whether obvious conformational changes exist between the substrate-free and substrate-bound cap-binding domain.
To study enzyme functionality, two haloalkane dehalogenase variants LinB32 and LinB70 carrying single-point and double-point mutations were constructed and crystallized in different crystallization conditions. Both LinB variants and their complexes with halogenated substrates diffracted to resolutions ranging from 1.6 to 2.8 Å.
β-Mannosidase from Aspergillus niger was crystallized in the presence of D-mannose and the crystal diffracted to 2.41 Å resolution. The crystal belonged to the space group P1 with unit-cell parameters a= 62.37, b = 69.73, c = 69.90 Å, α = 108.20, β = 101.51, γ = 103.20°.
The cellular pyrin domain-only protein 1 (cPOP1) was crystallized. The crystals were found to belong to the cubic space group P213 with unit-cell parameters a = b = c = 94.12 Å, α = β = γ = 90.00°. The crystals were obtained at 293 K and diffracted to a resolution of 3.6 Å.
The crystallization of three recombinant mutants of Vaccinia virus uracil DNA glycosylase is reported.
Crystals of the complex between the Fab fragment of a human anti-alpha toxin antibody and alpha toxin have been obtained. Diffraction data sets were collected to 2.56 Å resolution.
N-Acetylneuraminate lyase, an enzyme involved in the bacterial uptake and metabolism of sialic acid, is a promising target for antibiotic development against pathogenic bacteria. Here, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of N-acetylneuraminate lyase from methicillin-resistant S. aureus to 1.70 Å resolution are reported.
The expression, purification, crystallization, preliminary X-ray diffraction and molecular-replacement studies on C. parapsilosis carbonyl reductase are reported.
Diffraction data to 1.94 Å limiting resolution were collected from a crystal of selenomethionine-substituted LECT2.
The wild-type protein and four active-site mutants of xylanase II from Trichoderma reesei that catalyzes the hydrolysis of glycosidic bonds in xylan have successfully been crystallized. The crystallization of several structures including ligand-free and protein ligand complexes containing the substrate (xylohexaose) or product (xylotriose) are detailed.
Crystals of an S84D/S86D/S88D triple mutant of the casein kinase 2 interacting protein-1 (CKIP-1) pleckstrin homology domain were obtained and diffraction data were collected and processed to 1.7 Å resolution.
Here, Escherichia coli octaprenyl pyrophosphate synthase was expressed, purified and crystallized. The crystals were obtained by the sitting-drop vapour-diffusion method and diffracted to 2.2 Å resolution.
Peptidyl-tRNA hydrolase from T. thermophilus has been expressed, purified and crystallized. The crystals diffracted X-rays to atomic resolution (beyond 1.0 Å resolution).
laboratory communications
A simple fluorescence-based calibration method that can be used to monitor the precision and accuracy of any liquid-handling nano-dispenser device is presented.