Crystallization and preliminary X-ray analysis of human thioredoxin peroxidase-B from red blood cells.

Journal ArticleResearch Support, Non-U.S. Gov'tTwo different crystal forms of human thioredoxin peroxidase-B have been grown by vapour diffusion using polyethylene glycol 400 as a precipitant. Monoclinic P21 crystals were grown from freshly purified protein, whilst orthorhombic P212121 crystals were grown from purified protein that had been stored in ammonium sulfate, but otherwise under the same conditions. The diffraction from both crystal forms was observed to extend to beyond 2.0 A resolution using synchrotron radiation. Complete native data sets to 1.8 and 3. 7 A have been collected from the monoclinic and orthorhombic crystals, respectively.EU (HCMP Access to Large Scale Facilities grant

Crystallization and preliminary X-ray diffraction studies of the oxygenating subunit of 3,6-diketocamphane monooxygenase from Pseudomonas putida.

Journal ArticleResearch Support, Non-U.S. Gov'tThe oxygenating constituent of the 3,6-diketocamphane monooxygenase isozyme from Pseudomonas putida NCIMB 10007 has been crystallized under two different conditions. Crystals were initially grown from polyethylene glycol (PEG) 8000 and sodium acetate using the vapour-phase diffusion method. The crystals were of orthorhombic P212121 space group, with cell dimensions a = 55.8, b = 94.5 and c = 163.7 A and diffracted to 2.8 A resolution. More recently, improved crystals, which diffracted beyond 2 A, have been grown from ammonium sulfate. These crystals also belong to the orthorhombic P212121 space group, with cell dimensions of a = 54.6, b = 93.2 and c = 154. 1 A. A full native data set to 2.5 A resolution has been collected from the ammonium sulfate grown crystals.BBSR

Phenomenological analysis of the CLAS data on double charged pion photo and electro- production

First comprehensive data on the evolution of nucleon resonance photocouplings with photon virtuality Q^2 are presented for excited proton states in the mass range from 1.4 to 2.0 GeV. N^* photocouplings were determined in phenomenological analysis of CLAS data on 2 pi photo and electroproduction within the framework of the JLAB-MSU phenomenological model.Comment: 10 pages, 7 figures (encapsulated postscript

The Fasciola hepatica thioredoxin: High resolution structure reveals two oxidation states.

addresses: Henry Wellcome Building for Biocatalysis, School of Biosciences, University of Exeter, Stocker Road, Exeter EX4 4QD, UK.types: Comparative Study; Journal Article; Research Support, Non-U.S. Gov'tCopyright © 2008 Elsevier. NOTICE: this is the author’s version of a work that was accepted for publication in Molecular and Biochemical Parasitology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Molecular and Biochemical Parasitology, 2008, Vol. 161, Issue 1, pp. 44 – 48 DOI: 10.1016/j.molbiopara.2008.06.009The Fasciola hepatica thioredoxin protein structure has been determined to 1.45A resolution. This is the first example of a single crystal structure to show the active site cysteine residues in both the reduced and disulfide oxidised form. Consistent with this observation the process of oxidation appears to require very little rearrangement of the surrounding protein structure. The F. hepatica thioredoxin structure has been compared to other thioredoxin protein structures already known and is found to be highly conserved. The F. hepatica protein is most similar to that of the thioredoxin from its human and animal hosts but it resembles other parasitic thioredoxins with regard to having no additional cysteine residues and is therefore not regulated by transient disulfide bond formation as proposed for thioredoxins from higher eukaryotic species

Nucleon Resonance Structure from CLAS and CLAS12 Experiments

The recent results on the photo- and electrocouplings extraction from the reaction of one- and two-pion photo- and electroproduction off protons in the resonances region are presented. The production of two charged pions is of particular importance for evaluation of the photocouplings for the Δ(1620)12 −, Δ(1700)32 −

Dp breakup reaction investigation using polarized and unpolarized deuteron beam

Deuteron Spin Structure (DSS) collaboration program is aimed on few nucleon correlations investigation using unpolarized and polarized deuteron beam at intermediate energy range. Data of the dp breakup reaction have been obtained at energy range from 300 - 500 MeV of deuteron energy for various detector configurations in region where few nucleon correlations and relativistic effects can play significant rol

History-sensitive accumulation rules for life-time prediction under variable loading

This is the post-print version of the article. The official published version can be obtained from the link below - Copyright @ 2011 SpringerA general form of temporal strength conditions under variable creep loading is employed to formulate several new phenomenological accumulation rules based on the constant-loading durability diagram. Unlike the well-known Robinson rule of linear accumulation of partial life-times, the new rules allow to describe the life-time sensibility to the load sequence, observed in experiments. Comparison of the new rules with experimental data shows that they fit the data much more accurately than the Robinson rule

Substrate specificity of branched chain amino acid aminotransferases: The substitution of glycine to serine in the active site determines the substrate specificity for α-ketoglutarate

A branched chain aminotransferase from Thermoproteus tenax has been identi fi ed, cloned, over-expressed and biochemically characterised. A molecular modelling approach has been used to predict the 3D structure allowing its comparison with other related enzymes. This enzyme has high similarity to a previously characterised aminotransferase from Thermoproteus uzoniensis however its substrate speci fi city shows key differences towards the substrate α -ketoglutarate. Examination of the active sites of the two related enzymes reveals a single amino acid substitution of a glycine residue to a serine residue which could be responsible for this difference. When Gly104 in T. tenax was mutated to a serine residue and the resultant enzyme characterised, this single amino acid change resulted in a dramatic reduction in activity towards α - ketoglutarate with an 18-fold reduction in Vmax and a 20-fold Km increase, resulting in a 370-fold lower catalytic ef fi ciency. Structural comparisons between the two related Thermoproteus enzymes and another branched chain aminotransferase from Geoglobus acetivorans has revealed that the serine residue affects the fl exibility of a key loop involved in catalysis. This subtle difference has provided further insight into our understanding of the substrate speci fi city of these industrially important enzymes