5 resultados para 92-599B

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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The oxygenation of human Hb (HbA) demands a three state model: two deoxy states To and Tx, free and complexed with anions respectively, and an oxy R state. The regulation between these states is modulated by the presence of anions, such as chloride, that binds to T state. The b inding if chloride, however, remains controversial. The aim of this work is the study of arginines 92a (a1ß2 interface) and 141a (C-terminal) as chloride binding sites. To investigate that, we have studied 92 and 141 site directed mutant species: natural mutants Hb J-Cape-Town (R92Q), desArg (R141Δ), Chesapeake (R92L), and the constructed Chesapeake desArg (R92L,141Δ). We expressed Hbs in Escherichia coli and purified. Through oxygen binding curves we measured affinity and cooperativity, in function of water effect and Bohr effect in presence and absence of chloride. Structural features were obtained through 1H NMR spectroscopy Oxygen binding properties and Bohr effect measured indicated a higher affinity and lower cooperativity in absence and presence of chloride for all mutants. Structural changes represent functional aspects of mutant Hbs, such as a significant rise in affinity or a change in cooperativity. Water activity studies conducted as a function of chloride concentration showed that the only Hb desArg follows the thre state model. The other mutant Hbs do not exhibit the Tx state, a fact confirmed by the number of water molecules bound to each Hb during the deoxy-oxy transition. This behavior suggests that the Arginine 92 site could be responsible for chloride binding to Hb, since oxygenation of 92 mutant Hbs cannot be adjusted by the three state model. However, Bohr effect showed that all mutant Hbs released~1 proton in chloride presence, different from HbA that releases ~2, suggesting a role for 141 arginine in the tertiary and quaternary Bohr effect.

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This article discusses the controversy surrounding the winning project of the competition organized by the Brazilian Ministry of Foreign Affairs for the Brazilian pavilion at the Expo'92 in Seville, Spain; the views of architecture critics made at the time of the results, and, their implications for Brazilian architecture. At the beginning of the 90s, this competition was a kind of confrontation of ideas, between the architects who were in favour of a renewal of architecture and those who defended the resumption of national architectural traditions, buildings with large spans, constructed in reinforced concrete. These architects were the heirs of the so called Paulista architecture, which was characterised by the work undertaken from the 60's by important architects such as Vilanova Artigas and Paulo Mendes da Rocha. The modern references adopted by the winning project, from the architects Angelo Bucci, Alvaro Puntoni and José Oswaldo Vilela, sparked controversies because of the difficulty of resuming the teachings of the old modern masters when faced with new times. These controversies were related to the end of the military dictatorship in Brazil and the process of opening markets, and by the relevance of a re-evaluation of the so-called Paulista architecture.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)