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Resolution In-house (resolution + in-house)
Selected AbstractsStructure of a family 3b, carbohydrate-binding module from the Cel9V glycoside hydrolase from Clostridium thermocellum: structural diversity and implications for carbohydrate bindingACTA CRYSTALLOGRAPHICA SECTION D, Issue 1 2010Svetlana Petkun Family 3 carbohydrate-binding modules (CBM3s) are associated with both cellulosomal scaffoldins and family 9 glycoside hydrolases (GH9s), which are multi-modular enzymes that act on cellulosic substrates. CBM3s bind cellulose. X-ray crystal structures of these modules have established an accepted cellulose-binding mechanism based on stacking interactions between the sugar rings of cellulose and a planar array of aromatic residues located on the CBM3 surface. These planar-strip residues are generally highly conserved, although some CBM3 sequences lack one or more of these residues. In particular, CBM3b, from Clostridium thermocellum Cel9V exhibits such sequence changes and fails to bind cellulosic substrates. A crystallographic investigation of CBM3b, has been initiated in order to understand the structural reason(s) for this inability. CBM3b, crystallized in space group C2221 (diffraction was obtained to 2.0,Å resolution in-house) with three independent molecules in the asymmetric unit and in space group P41212 (diffraction was obtained to 1.79,Å resolution in-house and to 1.30,Å resolution at a synchrotron) with one molecule in the asymmetric unit. The molecular structure of Cel9V CBM3b, revealed that in addition to the loss of several cellulose-binding residues in the planar strip, changes in the backbone create a surface `hump' which could interfere with the formation of cellulose,protein surface interactions and thus prevent binding to crystalline cellulose. [source] Crystallization and preliminary X-ray analysis of inorganic pyrophosphatase from the hyperthermophilic archaeon Pyrococcus horikoshii OT3ACTA CRYSTALLOGRAPHICA SECTION D, Issue 3 2004Binbin Liu Inorganic pyrophosphatase (PPase; EC 3.6.1.1) from the hyperthermophile Pyrococcus horikoshii was crystallized by the hanging-drop vapour-diffusion method at pH 5.0 using polyethyleneglycol 4000 as the precipitant. The crystal belongs to space group P21212, with unit-cell parameters a = 71.7, b = 86.5, c = 92.5,Å, , = , = , = 90°. There are two molecules in the asymmetric unit. The crystals were stable during exposure to X-rays and a full set of X-ray diffraction data was collected to 2.7,Å resolution in-house. [source] Cloning, purification, crystallization and X-ray crystallographic analysis of Ignicoccus hospitalis neelaredoxinACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 5 2010Filipa G. Pinho Superoxide reductases (SORs) are metalloproteins which constitute the most recently identified oxygen-detoxification system in anaerobic and microaerobic bacteria and archaea. SORs are involved in scavenging superoxide radicals from the cell by catalyzing the reduction of superoxide () to hydrogen peroxide and are characterized by a catalytic nonhaem iron centre coordinated by four histidine ligands and one cysteine ligand. Ignicoccus hospitalis, a hyperthermophilic crenarchaeon, is known to have a neelaredoxin-type SOR that keeps toxic oxygen species levels under control. Blue crystals of recombinant I. hospitalis oxidized neelaredoxin (14.1,kDa, 124 residues) were obtained. These crystals diffracted to 2.4,Å resolution in-house at room temperature and belonged to the hexagonal space group P6222 or P6422, with unit-cell parameters a = b = 108, c = 64,Å. Cell-content analysis indicated the presence of one monomer in the asymmetric unit. [source] Crystallization and crystallographic analysis of the apo form of the orange protein (ORP) from Desulfovibrio gigasACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2009Shabir Najmudin The orange-coloured protein (ORP) from Desulfovibrio gigas is a 12,kDa protein that contains a novel mixed-metal sulfide cluster of the type [S2MoS2CuS2MoS2]. Diffracting crystals of the apo form of ORP have been obtained. Data have been collected for the apo form of ORP to 2.25,Å resolution in-house and to beyond 2.0,Å resolution at ESRF, Grenoble. The crystals belonged to a trigonal space group, with unit-cell parameters a = 43, b = 43, c = 106,Å. [source] Crystallization and preliminary X-ray analysis of carnein, a serine protease from Ipomoea carneaACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 4 2009Ashok Kumar Patel Carnein is an 80,kDa subtilisin-like serine protease from the latex of the plant Ipomoea carnea which displays an exceptional resistance to chemical and thermal denaturation. In order to obtain the first crystal structure of a plant subtilisin and to gain insight into the structural determinants underlying its remarkable stability, carnein was isolated from I. carnea latex, purified and crystallized by the hanging-drop vapour-diffusion method. A data set was collected to 2.0,Å resolution in-house from a single crystal at 110,K. The crystals belonged to the trigonal space group P3121 or P3221, with unit-cell parameters a = b = 126.9, c = 84.6,Å, , = , = 90, , = 120°. Assuming the presence of one molecule per asymmetric unit, the Matthews coefficient is 2.46,Å3,Da,1, corresponding to a solvent content of 50%. Structure determination of the enzyme is in progress. [source] Expression, purification, crystallization and preliminary crystallographic study of the carboxyl-terminal domain of the human voltage-gated proton channel Hv1ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 3 2009Shu Jie Li The voltage-gated proton channel Hv1 is essential to proton permeation and contains a voltage-sensor domain without a pore domain. It contains three predicted domains: an N-terminal acid and proline-rich domain, a transmembrane voltage-sensor domain and a C-terminal domain that is responsible for the dimeric architecture of Hv1. Here, the C-terminal domain of the human voltage-gated proton channel Hv1 (C-Hv1) was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method. The crystals have a tetragonal form and diffraction data were collected to 2.5,Å resolution in-house. The crystal belongs to space group P41212, with unit-cell parameters a = b = 37.76, c = 137.52,Å. Structural determination of C-Hv1 is in progress. [source] Crystallization and X-ray diffraction analysis of the DNA-remodelling protein DnaD from Bacillus subtilisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 2 2007Sabine Schneider The DnaD protein is an essential component of the chromosome-replication machinery of the Gram-positive bacterium Bacillus subtilis and is part of the primosomal cascade that ultimately loads the replicative ring helicase DnaC onto DNA. Moreover, DnaD is a global regulator of DNA architecture, as it forms higher order nucleoprotein structures in order to open supercoiled DNA. Here, the crystallization and preliminary X-ray diffraction analysis of the two domains of DnaD from B. subtilis are reported. Crystals of the N-terminal domain are trigonal, with either P3121 or P3221 space-group symmetry, and diffracted X-rays to 2.0,Å resolution; crystals of the C-terminal domain are hexagonal, with space group P61 or P65, and diffracted X-rays to 2.9,Å resolution in-house. Determination of the structure of the DnaD domains will provide insight into how remodelling of the nucleoid is associated with priming of replication in the model Gram-positive organism B. subtilis. [source] | ||||||||||