Structure of BthA-I complexed with p-bromophenacyl bromide: possible correlations with lack of pharmacological activity

The crystal structure of an acidic phospholipase A(2) isolated from Bothrops jararacussu venom (BthA-I) chemically modified with p-bromophenacyl bromide (BPB) has been determined at 1.85 angstrom resolution. The catalytic, platelet-aggregation inhibition, anticoagulant and hypotensive activities of BthA-I are abolished by ligand binding. Electron-density maps permitted unambiguous identification of inhibitor covalently bound to His48 in the substrate-binding cleft. The BthA-I-BPB complex contains three structural regions that are modified after inhibitor binding: the Ca2+-binding loop, ss-wing and C-terminal regions. Comparison of BthA-I-BPB with two other BPB-inhibited PLA(2) structures suggests that in the absence of Na+ ions at the Ca2+- binding loop, this loop and other regions of the PLA(2)s undergo structural changes. The BthA-I-BPB structure reveals a novel oligomeric conformation. This conformation is more energetically and conformationally stable than the native structure and the abolition of pharmacological activities by the ligand may be related to the oligomeric structural changes. A residue of the `pancreatic' loop (Lys69), which is usually attributed as providing the anticoagulant effect, is in the dimeric interface of BthA-I-BPB, leading to a new hypothesis regarding the abolition of this activity by BPB.

Laurdan Spectrum Decomposition as a Tool for the Analysis of Surface Bilayer Structure and Polarity: a Study with DMPG, Peptides and Cholesterol

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

Foraging by the omnivorous crab Pachygrapsus transversus affects the structure of assemblages on sub-tropical rocky shores

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

Population structure and reproductive period of the tropical hermit crab Calcinus tibicen (Decapoda : Diogenidae) in the region of Ubatuba, São Paulo, Brazil

The population of the hermit crab Calcinus tibicen in the Ubatuba region was studied with emphasis on seasonal abundance, seasonal size-frequency distribution, sex-ratio, and reproductive period based on the percentage of ovigerous females. The animals were collected at 2-month intervals for 2 consecutive years (from January 1993 to November 1994). Total mean animal size in shield length was 5.14 +/- 1.23 mm for males, 4.23 +/- 0.79 mm for females, and 4.53 +/- 0.60 for ovigerous females. A discontinuity in reproduction was observed, with an absence of ovigerous females in July (winter) and a high incidence from September to May (spring to autumn). Calcinus tibicen is sexually dimorphic in relation to size, with a higher growth rare in males. The sex-ratio pattern changes with season, indicating that it is more a function of season or habitat than of size.

Crystallographic structure of PNP from Mycobacterium tuberculosis at 1.9 angstrom resolution

Even being a bacterial purine nucleoside phosphorylase (PNP), which normally shows hexameric folding, the Mycobacterium tuberculosis PNP (MtPNP) resembles the mammalian trimeric structure. The crystal structure of the MtPNP apoenzyme was solved at 1.9 Angstrom resolution. The present work describes the first structure of MtPNP in complex with phosphate. In order to develop new insights into the rational drug design, conformational changes were profoundly analyzed and discussed. Comparisons over the binding sites were specially studied to improve the discussion about the selectivity of potential new drugs. (C) 2004 Elsevier B.V. All rights reserved.

Structure of chorismate synthase from Mycobacterium tuberculosis

In bacteria, fungi, plants, and apicomplexan parasites, the aromatics compounds, such as aromatics amino acids, are synthesized through seven enzymes from the shikimate pathway, which are absent in mammals. The absence of this pathway in mammals make them potential targets for development of new therapy against infectious diseases, such as tuberculosis, which is the world's second commonest cause of death from infectious disease. The last enzyme of shikimate pathway is the chorismate synthase (CS), which is responsible for conversion of the 5-enolpyruvylshikimate-3-phosphate to chorismate. Here, we report the crystallographic structure of CS from Mycobacterium tuberculosis (MtCS) at 2.65 angstrom resolution. The MtCS structure is similar to other CS structures, presenting beta-alpha-beta sandwich structural topology, in which each monomer of MtCS consists of a central helical core. The MtCS can be described as a tetramer formed by a dimer of dimers. However, analytical ultracentrifugation studies suggest the MtCS is a dimer with a more asymmetric shape than observed on the crystallographic dimer and the existence of a low equilibrium between dimer and tetramer. Our results suggest that the MtCS oligomerization is concentration dependent and some conformational changes must be involved on that event. (c) 2005 Elsevier B.V. All rights reserved.

Dynamics of glyphosate-induced conformational changes of Mycobacterium tuberculosis 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19) determined by hydrogen-deuterium exchange and electrospray mass spectrometry

The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) catalyzes the reaction between shikimate 3-phosphate and phosphoenolpyruvate to form 5-enolpyruvylshikimate 3-phosphate, an intermediate in the shikimate pathway, which leads to the biosynthesis of aromatic amino acids. EPSPS exists in an open conformation in the absence of substrates and/or inhibitors and in a closed conformation when bound to the substrate and/or inhibitor. In the present report, the H/D exchange properties of EPSPS from Mycobacterium tuberculosis (Mt) were investigated for both enzyme conformations using ESI mass spectrometry and circular dichroism (CD). When the conformational changes identified by H/D exchanges were mapped on the 3-D structure, it was observed that the apoenzyme underwent extensive conformational changes due to glyphosate complexation, characterized by an increase in the content of alpha-helices from 40% to 57%, while the beta-sheet content decreased from 30% to 23%. These results indicate that the enzyme underwent a series of rearrangements of its secondary structure that were accompanied by a large decrease in solvent access to many different regions of the protein. This was attributed to the compaction of 71% of alpha-helices and 57% of beta-sheets as a consequence of glyphosate binding to the enzyme. Apparently, MtEPSPS undergoes a series of inhibitor-induced conformational changes, which seem to have caused synergistic effects in preventing solvent access to the core of molecule, especially in the cleft region. This may be part of the mechanism of inhibition of the enzyme, which is required to prevent the hydration of the substrate binding site and also to induce the cleft closure to avoid entrance of the substrates.

Ultra-structure and histochemistry of digestive cells of Podisus nigrispinus (Hemiptera: Pentatomidae) fed with prey reared on bt-cotton

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The significance of changes in Mytella falcata (Orbigny, 1842) gill filaments chronically exposed to polluted environments

The impact of pollutants in an organism can be observed by changes in functional complexity at different levels. Bivalve gills are suitable for histopathological analysis because of their structure and function. This study aimed at examining the morphology of Mytella falcata gill filaments from three sites in the Santos estuary (São Paulo, Brazil) with different levels of environmental degradation to identify possible changes in gill structure and discuss the significance of these alterations. For this purpose, histological, histochemical and ultrastructural techniques were used. The filaments of animals from site A (less impacted site) were intact, while in sites B and C, pathological changes were observed, such as: detachment of the epithelium in the intermediate zone, morphological changes of this epithelium, inflammatory process, increase in the number of mucous cells and cell turnover processes. These results suggest that the related changes are an attempt to prevent the entrance of pollutants through gill filaments into the entire organism and that cell turnover is the final way to compensate cell injury. (C) 2008 Elsevier Ltd. All rights reserved

Structure and function of the foregut and salivary glands of the synanthropic diplopod Urostreptus atrobrunneus (Spirostreptidae)

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

Short-term changes caused by fire and mowing in Brazilian Campos grasslands with different long-term fire histories

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Invasive Africanized honeybees change the structure of native pollination networks in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Structure of the posthepatic septum and its influence on visceral topology in the tegu lizard, Tupinambis merianae (Teiidae : Reptilia)

The posthepatic septum (PHS) divides the body cavity of Tupinambis merianae into two parts: the cranial one containing the lungs and liver and the caudal one containing the remaining viscera. The PHS is composed of layers of collagenous fibers and bundles of smooth muscle, neither of which show systematic orientation, as well as isolated blood vessels, lymphatic vessels, and nerves. Striated muscle of the abdominal wall does not invade the PHS. The contractions of the smooth muscles may stabilize the pleurohepatic cavity under conditions of elevated aerobic needs rather than supporting breathing on a breath-by-breath basis. Surgical removal of the PHS changes the anatomical arrangement of the viscera significantly, with stomach and intestine invading the former pleurohepatic cavity and reducing the space for the lungs, Thus, the PHS is essential to maintain the visceral topography in Tupitionibis. J. Morphol. 258:151-157, 2003. (C) 2003 Wiley-Liss. Inc.

Composition and structure of fish assemblage from Passa Cinco stream, Corumbataí river sub-basin, SP, Brazil

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

Changes in ichthyofauna composition along a gradient from clearwaters to blackwaters in coastal streams of Atlantic forest (southeastern Brazil) in relation to environmental variables

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