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October 2009 issue
Cover illustration: Use of the MAD triangle B3C with bovine trypsin (Beck et al., p. 1068) as described in the first article in the new Laboratory Communications category.
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structural communications
The crystal structures of truncated forms of the Streptococcus pyogenes phage-encoded hyaluronate lyases HylP2 and HylP3 have been determined by molecular replacement to 1.6 and 1.9 Å resolution, respectively.
Monovalent cation-binding sites in GroEL were identified utilizing the anomalous scattering signal from Tl+.
PDB reference: GroEL, 3e76, r3e76sf
The crystal structure of Rv2714, a protein of unknown function from M. tuberculosis, has been determined at 2.6 Å resolution using single-wavelength anomalous diffraction methods.
PDB reference: Rv2714, 2wam, r2wamsf
The first crystal structure of 4-methyl-5-β-hydroxyethylthiazole kinase from an archaeon (P. horikoshii OT3) has been determined at 1.85 Å resolution. Comparative analyses of sequences and structures and modelling studies are presented.
PDB reference: 4-methyl-5-β-hydroxyethylthiazole kinase, 3hpd, r3hpdsf
The crystal structure of S. aureus phosphopantetheine adenylyltransferase in complex with 3′-phosphoadenosine 5′-phosphosulfate has been determined and reveals a new ligand-binding mode.
PDB reference: PAPS-bound S. aureus PPAT, 3f3m, r3f3msf
The crystal structure of human carbonic anhydrase II (CA II) complexed with acetazolamide (AZM) has been determined at 1.1 Å resolution. The co-binding of AZM and glycerol in the active site demonstrate that an isozyme specific CA inhibitor may be developed.
PDB reference: carbonic anhydrase II, 3hs4, 3hs4sf
crystallization communications
The major outer membrane protein PorB from N. meningitidis was crystallized in three crystal forms; the best X-ray diffraction data were collected to 2.3 Å resolution.
The X-ray diffraction and preliminary phasing of the putative transcriptional regulator Bxe_C0898 from B. xenovorans LB400 are reported.
The crystallization and preliminary crystallographic analysis of a para-nitrophenol 4-monooxygenase PnpA from Pseudomonas putida DLL-E4 are presented.
A variant of StHsp14.0, a small heat-shock protein from S. tokodaii, has been crystallized. Heavy-atom derivative crystals could be prepared by the cocrystallization method.
Crotoxin B is a basic phospholipase A2 found in the venom of C. durissus collilineatus and is one of the subunits which constitute the crotoxin. Here, we present the crystallization, X-ray diffraction data collection and molecular-replacement solution of the dimeric complex formed by two crotoxin B isoforms.
Mannosyl-3-phosphoglycerate synthase (MpgS) is a key enzyme in the biosynthesis of MG. Here, the purification, crystallization and preliminary crystallographic characterization of apo MpgS from Thermus thermophilus HB27 are reported.
The hatching enzyme of zebrafish, ZHE1, was expressed, purified and crystallized using the hanging-drop vapour-diffusion method. The crystal belonged to space group P212121 and diffracted X-rays to a resolution of 1.14 Å.
Crystals of recombinant RpfF from S. maltophilia are tetragonal, space group P41212 or P43212, with unit-cell parameters a = 148.51, c = 122.82 Å, and diffract to 2.25 Å resolution.
The VanG D-alanine:D-serine ligase was crystallized in complex with ADP and diffraction data were collected at 2.35 Å resolution.
Parakeet (Psittacula krameri) haemoglobin has been purified and crystallized under low salt buffered conditions. Preliminary analysis of the crystal that belonged to monoclinic system (C2) is reported.
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Algal Chlorella virus IL-3A deoxyuridine triphosphatase and its Glu81Ser plus Thr84Arg-mutated derivative, Mu-22, were crystallized using the hanging-drop vapor-diffusion method. Glycyl-seryl-tagged dUTPases yielded cubic and hexagonal crystals for IL-3A and Mu-22, respectively.
P. aeruginosa RocR, an EAL-domain protein, has been expressed in E. coli, purified and crystallized. An X-ray diffraction data set was collected to a resolution of 2.50 Å.
The O-methyltransferase (OMT) from the Anabaena PCC 7120 has been overexpressed in a soluble form in E. coli, purified and crystallized. The crystals belonged to space group C2221 and diffracted to 2.4 Å resolution.
Various N-terminally truncated forms of the S-layer protein SbsC were crystallized and the form rSbsC(755–1099), which was obtained by unintentional in situ proteolysis, was used for preliminary structure determination.
Two shikimate-pathway enzymes from M. tuberculosis, the intracellular chorismate mutase (MtCM) and DAHP synthase (MtDS), were produced recombinantly and purified. MtCM was crystallized alone and in complex with MtDS and analyzed by X-ray diffraction.
A nitric oxide-inducible lactate dehydrogenase from S. aureus, Sa-LDH-1, has been cloned, expressed, purified and crystallized in space group C2, with unit-cell parameters a = 131.4, b = 74.4, c = 103.2 Å, β = 133.4°.
An essential protein from the plant pathogen X. campestris pv. campestris (Xcc) that contains a noncanonical PilZ signature motif yet is critical for Xcc pathogenicity has been overexpressed in E. coli, purified and crystallized. The crystals diffracted to a resolution of 2.1 Å.
The crystallization of the putative MIF4G domain of Paip1 is described. The crystals belonged to the monoclinic space group P21 and diffracted X-rays to beyond 2.2 Å resolution.
In this study, two forms of MJ0754 from the archaeon M. jannaschii were overexpressed and crystallized. The crystal of MJ0754 belonged to the hexagonal space group P61 and diffracted to 3.1 Å resolution, while the crystal of MJ0754t belonged to the orthogonal space group C2221 and diffracted to 1.3 Å resolution using synchrotron radiation.
laboratory communications
The handling of the phasing tools I3C and B3C is described, emphasizing practical aspects such as the preparation of solutions and incorporation of the compounds into protein crystals.
PDB reference: bovine trypsin crystallized with B3C, 3iti, r3itisf
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