The structure of a catalytically active subdomain of the NanA sialidase from S. pneumoniae is reported to a resolution of 2.5 Å. The complex with the inhibitor Neu5Ac2en identifies the key catalytic residues and provides a platform for structure-based development of specific inhibitors.
The structure of the catalytic subunit of M. jannaschii aspartate transcarbamoylase has been determined in space group P212121 using synchrotron data to a resolution of 3.0 Å and was refined to a final Rwork and Rfree of 0.215 and 0.269, respectively.
Acidic nondenaturing PAGE revealed inhomogeneity of the MASP-1 catalytic region caused by deamidation as the reason behind previous unsuccessful crystallization attempts. Monitoring the separation of the various species by acidic nondenaturing PAGE during purification helped to obtain pure protein, which was subsequently crystallized.
Histamine dehydrogenase from Nocardioides simplex has been expressed, purified and crystallized with full incorporation of 6-S-cysteinyl-FMN. Diffraction data have been collected to 2.7 Å resolution; the crystals belonged to the orthorhombic space group P212121.
Crystallization of the proposed FAD-containing ferri-siderophore reductase protein YqjH from E. coli has been performed and the structure has been determined to 3.0 Å resolution. The structure shows similarity to another proposed siderophore-interacting protein from S. putrefaciens and weak similarity to members of the NAD(P)H:flavin oxidoreductase superfamily.
The full length and the regulatory domain of the LysR-type transcriptional regulator CrgA have been crystallized. Diffraction data were collected from two crystal forms of full-length CrgA to 3.0 and 3.8 Å resolution, respectively. Crystals of the selenomethionine derivative of the C-terminal regulatory domain of CrgA diffracted to 2.3 Å resolution.
N-Acetylglucosamine 1-phosphate uridyltransferase (GlmU) from M. tuberculosis H37Rv has been crystallized and preliminary X-ray crystallographic analysis has been performed. GlmU is a bi-domained bifunctional enzyme that is involved in the biosynthesis of UDP-N-acetylglucosamine, a precursor in peptidoglycan biosynthesis in M. tuberculosis.
The cloning, expression, purification and crystallization of the complex of the second and third regulatory subunits of human Pol δ are reported. The crystals were characterized and an X-ray diffraction data set was collected to a resolution of 3 Å.
The L-threonine dehydrogenase from T. kodakaraensis KOD1 has been crystallized in the tetragonal space group P43212, with unit-cell parameters a = b = 124.5, c = 271.1 Å. Diffraction data were collected to 2.6 Å resolution and preliminary analysis indicates that there are four molecules in the crystallographic asymmetric unit.
A truncated variant of the human RuvBL1–RuvBL2 complex was cloned, expressed, purified and crystallised. Synchrotron diffraction data to 4 Å resolution were used to carry out a preliminary crystallographic analysis of the complex.
Succinate:ubiquinone oxidoreductase was solubilized and purified from E. coli inner membranes using several different detergents and the phospholipid content of each preparation was analyzed. The preparation with the lowest phospholipid content crystallized in two different crystal forms using detergent mixtures composed of n-alkyl-oligoethylene glycol monoether and n-alkyl-maltoside.
M. tuberculosis tetrahydrodipicolinate-N-succinyltransferase, the enzyme that catalyses the fifth reaction step of the lysine-biosynthesis pathway, has been cloned, expressed, purified and crystallized.
Recombinant RuBisCO chaperone from T. elongatus, TeRbcX, was aggregated and could not be crystallized. Mutations of an unusual Cys residue in the TeRbcX sequence yielded much better behaved proteins that could rapidly be crystallized.