Crystallization and preliminary X-ray diffraction analysis of the catalytic subunit of ADP-glucose pyrophosphorylase from potato tuber

ADP-glucose pyrophosphorylase is the key regulatory enzyme in the biosynthesis of starch in plants and glycogen in bacteria. The enzyme from potato tuber is comprised of a regulatory subunit and a catalytic subunit and is present as a heterotetramer (alpha(2)beta(2)) the catalytic subunit from potato tuber (50 kDa) was crystallized in four different forms, two of which are suitable for structural studies. A tetragonal crystal form obtained in the presence of the substrate analog Cr-ATP diffracted to 2.2 Angstrom and belongs to space group P4(1) (or its enantiomorph), with unit-cell parameters a = b = 110.57, c = 190.14 Angstrom. A second crystal form obtained diffracted to 2.8 Angstrom and belongs to space group PZ, with unit-eel parameters a = 80.06, b = 138.84, c = 92.20 Angstrom, beta = 112.40 degrees. As this protein displays no significant homology to any currently known protein structure, a search for heavy-atom derivatives has been initiated.

Crystallization and preliminary X-ray crystallographic analysis of the heterodimeric crotoxin complex and the isolated subunits crotapotin and phospholipase A(2)

Crotoxin, a potent neurotoxin from the venom of the South American rattlesnake Crotalus durissus terrificus, exists as a heterodimer formed between a phospholipase A(2) and a catalytically inactive acidic phospholipase A(2) analogue (crotapotin). Large single crystals of the crotoxin complex and of the isolated subunits have been obtained. The crotoxin complex crystal belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 38.2, b = 68.7, c = 84.2 angstrom, and diffracted to 1.75 angstrom resolution. The crystal of the phospholipase A(2) domain belongs to the hexagonal space group P6(1)22 (or its enantiomorph P6(5)22), with unit-cell parameters a = b = 38.7, c = 286.7 angstrom, and diffracted to 2.6 angstrom resolution. The crotapotin crystal diffracted to 2.3 angstrom resolution; however, the highly diffuse diffraction pattern did not permit unambiguous assignment of the unit-cell parameters.

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.

Crystallization and preliminary X-ray diffraction analysis of a class II phospholipase D from Loxosceles intermedia venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Production, purification, crystallization and preliminary X-ray diffraction studies of the nucleoside diphosphate kinase b from Leishmania major

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Crystallization and preliminary X-ray diffraction analysis of an l-amino-acid oxidase from Bothrops jararacussu venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification, crystallization and preliminary X-ray diffraction analysis of crotamine, a myotoxic polypeptide from the Brazilian snake Crotalus durissus terrificus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Purification, crystallization and preliminary X-ray diffraction analysis of a class P-III metalloproteinase (BmMP-III) from the venom of Bothrops moojeni

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Crystallization, preliminary X-ray diffraction analysis and Patterson search of oxyhaemoglobin I from the wolf (Chrysocyon brachiurus)

Oxyhaemoglobin I isolated from the Brazilian wolf Chrysocyon brachiurus has been crystallized and X-ray diffraction data has been collected to 2.06 A resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P2(1)2(1)2(1) and preliminary structural analysis revealed the presence of one tetramer in the asymmetric unit. The structure was determined using standard molecular-replacement techniques and is currently being refined using maximum-likelihood protocols. This is the first haemoglobin isolated from a member of the Canidae family to be crystallized and it will provide further insights in the comparative biochemistry of vertebrate haemoglobins.

Structural modifications in stretch-induced crystallization in PVDF films as measured by small-angle X-ray scattering

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of partial crystallization on the mechanical properties and cytotoxicity of bioactive glass from the 3CaO center dot P2O5-SiO2-MgO system

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of annealing on the crystallization and roughness of PLZT thin films

Lead lanthanum zirconate titanate (PLZT) thin films with stoichiometry (9/65/35) were prepared by a dip-coating process using a polymeric organic solution. The solution viscosity was adjusted in the range of 15-56 cP. Silicon (100) substrates were previously cleaned and then immersed in the solution. The withdrawal speed of substrate from the solution was adjusted within a range of 5 to 20 mm/min. The coated substrates were thermally treated in the 450-700 degreesC temperature range. Surface roughness and crystallization of these films are strongly dependent on the annealing conditions. Infrared and X-ray diffraction data for PLZT powders heat-treated at 650 degreesC for 3 h show that the material is free of carbonate phases and crystalline. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Influence of oxygen flow on crystallization and morphology of LiNbO3 thin films

Polymeric precursor solution (Pechini method) was used to deposit LiNbO3 thin films by spin-coating on (100) silicon substrates. X-ray diffraction data of thin films showed that the increase of oxygen flow promotes a preferred orientation of (001) LiNbO3 planes parallel to the substrate surface. Surface roughness and grain size, observed by atomic force microscopy, change also with oxygen flow.

Influence of oxygen flow on crystallization and morphology of PLZT thin films

PLZT thin films were prepared by a dip coating process using Pechini's method, also known as polymeric precursor method. The PLZT solution was obtained from a mixture of the individual cation solutions and the process to prepare each solution is based on metallic citrate polymerization. The viscosity of the PLZT solution was adjusted at 40 cP while the ionic concentration was adjusted at 0.1 M. PLZT solutions were deposited on silicon (100) and platinum coated silicon (100) substrates with withdrawal speed at 5 mm/min. The coated substrates were thermally treated with a heating rate of 1 degreesC/min up to 300 degreesC and 5 degreesC/min up to 650 degreesC in order to obtain homogeneous and cracks free films. The influence of oxygen flow on crystallization and morphology of PLZT (9/65/35) thin film is discussed. (C) 2002 Elsevier B.V. Ltd and Techna S.r.l. All rights reserved.

Influence of thickness on crystallization and properties of LiNbO3 thin films

Thin films of lithium niobate were deposited on Pt/Ti/SiO2 (111) substrates by spin coating from the polymeric precursor method (Pechini process). Annealing in static air was performed at 500 degreesC for 3 h. The obtained films were characterized by X-ray diffraction and atomic force microscopy. The dielectric constant, dissipation factor and resistance were measured in frequency region from 10 Hz to 10 MHz and the hysteresis loop was obtained. The influence of number of layers on crystallization, morphology and properties of LiNbO3 thin films is discussed. (C) 2003 Elsevier B.V. All rights reserved.