Crystal structure of an acidic platelet aggregation inhibitor and hypotensive phospholipase A(2) in the monomeric and dimeric states: insights into its oligomeric state

Phospholipases A(2) belong to the superfamily of proteins which hydrolyzes the sn-2 acyl groups of membrane phospholipids to release arachidonic acid and lysophospholipids. An acidic phospholipase A(2) isolated from Bothrops juraracussu snake venom presents a high catalytic, platelet aggregation inhibition and hypotensive activities. This protein was crystallized in two oligomeric states: monomeric and dimeric. The crystal structures were solved at 1.79 and 1.90 Angstrom resolution, respectively, for the two states. It was identified a Na+ ion at the center of Ca2+-binding site of the monomeric form. A novel dimeric conformation with the active sites exposed to the solvent was observed. Conformational states of the molecule may be due to the physicochemical conditions used in the crystallization experiments. We suggest dimeric state is one found in vivo. (C) 2004 Elsevier B.V. All rights reserved.

The effect of acidic residues and amphipathicity on the lytic activities of mastoparan peptides studied by fluorescence and CD spectroscopy

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

Insights into metal ion binding in phospholipases A(2): ultra high-resolution crystal structures of an acidic phospholipase A(2) in the Ca2+ free and bound states

The electrophile Ca2+ is an essential multifunctional co-factor in the phospholipase A(2) mediated hydrolysis of phospholipids. Crystal structures of an acidic phospholipase A(2) from the venom of Bothrops jararacussu have been determined both in the Ca2+ free and bound states at 0.97 and 1.60 angstrom resolutions, respectively. In the Ca2+ bound state, the Ca2+ ion is penta-coordinated by a distorted pyramidal cage of oxygen and nitrogen atoms that is significantly different to that observed in structures of other Group I/II phospholipases A(2). In the absence of Ca2+, a water molecule occupies the position of the Ca2+ ion and the side chain of Asp49 and the calcium-binding loop adopts a different conformation. (c) 2005 Elsevier SAS. All rights reserved.

A stability transition at mildly acidic pH in the alpha-hemolysin (alpha-toxin) from Staphylococcus aureus

The effects of mildly acidic conditions on the free energy of unfolding (Delta G(u)(buff)) of the pore-forming alpha-hemolysin (alpha HL) from Staphylococcus aureus were assessed between pH 5.0 and 7.5 by measuring intrinsic tryptophan fluorescence, circular dichroism and elution time in size exclusion chromatography during urea denaturation, Decreasing the pH from 7.0 to 5.0 reduced the calculated Delta G(u)(buff) from 8.9 to 4.2 kcal moI(-1), which correlates with an increased rate of pore formation previously observed over the same pH range, It is proposed that the lowered surface pH of biological membranes reduces the stability of alpha HL thereby modulating the rate of pore formation. (C) 1999 Federation of European Biochemical Societies.

The Venomics of Bothrops alternatus is a Pool of Acidic Proteins with Predominant Hemorrhagic and Coagulopathic Activities

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

Efficiency of ethanol conversion induced by controlled modification of pore structure and acidic properties of alumina catalysts

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Electrochemical Oxidation of Hydroxylamine on Gold in Aqueous Acidic Electrolytes: An in Situ SERS Investigation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Oxidation of Hydroxylamine on Gold Electrodes in Mildly Acidic Aqueous Electrolytes: Electrochemical and In Situ Differential Reflectance Studies

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of calcium pre-rinse and fluoride dentifrice on remineralisation of artificially demineralised enamel and on the composition of the dental biofilm formed in situ

Objective: This in situ blind crossover study investigated the effect of calcium (Ca) rinse prior to the use fluoride (F) dentifrice on remineralisation of artificially demineralised enamel and on the composition of biofilm. Design: During four phases of 14 days, 10 volunteers wore appliances containing two artificially demineralised bovine enamel blocks. Three times a day, they rinsed with 10 mL, of Ca (150 mM) or placebo rinse (1 min). A slurry (1:3, w/v) of F (1030 ppm) or placebo dentifrice was dripped onto the blocks. During I min, the volunteers brushed their teeth with the respective dentifrice. The appliance was replaced into the mouth and the volunteers rinsed with water. The biofilm formed on the blocks was analysed for F and Ca. Enamel alterations were evaluated by the percentage of surface microhardness change (%SMHC), cross-sectional microhardness (% mineral volume) and alkali-soluble F analysis. Data were analysed by ANOVA (p < 0.05). Results: the use of the Ca pre-rinse before the F dentifrice produced a six- and four-fold increase in biofilm F and Ca concentrations, respectively. For enamel, the remineralisation was significantly improved by the Ca pre-rinse when compared to the other treatments. There was a significantly higher concentration of alkali-soluble F in enamel when the F dentifrice was used, but the Ca pre-rinse did not have any significant additive effect. Conclusions: According to our protocol, the Ca pre-rinse significantly increased biofilm F concentration and, regardless the use of F dentifrice, significantly enhanced the remineralisation of artificially demineralised enamel. (c) 2007 Elsevier Ltd. All rights reserved.

Correlation between acidic ninhydrin and HPLC methods to evaluate fraudulent addition of whey in milk

The acidic ninhydrin spectrophotometric method (ANSM) for quantitative determination of free and bound sialic acid of milk glycoprotein has been proved to be fast and efficient for routine detection of fraudulent addition of rennet whey to fluid milk. In this research the ANSM was compared with the high performance liquid chromatography (HPLC) method, internationally recommended for caseinomacropeptide (CMP) determination, which besides its high accuracy is more sophisticated and requires trained personnel. For several sample conditions (raw milk and milk with variable added amounts of rennet cheese whey), the methods showed an excellent linear correlation, with r = 0.981 when milk was deproteinized with a 120 g.L-1 final concentration of trichloroacetic acid (TCA) concentration. The best correlations could be seen with final concentrations of 100 g.L-1 and 80 g.L-1 TCA; respectively, r = 0.992 and 0.993.

Chromotropic acid-formaldehyde reaction in strongly acidic media. The role of dissolved oxygen and replacement of concentrated sulphuric acid

The procedure for formaldehyde analysis recommended by the National Institute for Occupational Safety and Health (NIOSH) is the Chromotropic acid spectrophotometric method, which is the one that uses concentrated sulphuric acid. In the present study the oxidation step associated with the aforementioned method for formaldehyde determination was investigated. Experimental evidence has been obtained indicating that when concentrated H2SO4 (18 mol l(-1)) is used (as in the NIOSH procedure) that acid is the oxidizing agent. on the other hand, oxidation through dissolved oxygen takes place when concentrated H2SO4 is replaced by concentrated hydrochloric (12 mol l(-1)) and phosphoric (14.7 mol l(-1)) acids as well as by diluted H2SO4 (9.4 mol l(-1)). Based on investigations concerning the oxidation step, a modified procedure was devised, in which the use of the potentially hazardous and corrosive concentrated H2SO4 was eliminated and advantageously replaced by a less harmful mixture of HCl and H2O2. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Neodymium biosorption from acidic solutions in batch system

Biosorption of neodymium in batch experiments took similar to 2 h to achieve the equilibrium biosorbent-metal for all microorganisms tested. The best biosorption coefficient at a constant pH value of 1.5 was obtained using the microalgae Monoraphidium sp. (1521 mg g(-1) cell), followed by Bakers' yeast (313 mg g(-1) cell), Penicillium sp. (178 mg g(-1) cell), and activated carbon (61 mg g(-1) cell). When compared to the biosorption of other metals, these results pointed out to the application of biosorption in neodymium recovery from acidic solutions. (C) 2000 Elsevier B.V. Ltd. All rights reserved.