13 resultados para aminopeptidase

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


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Pós-graduação em Microbiologia Agropecuária - FCAV

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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A variabilidade genética foi avaliada dentro e entre amostras de diferentes cultivares de amendoim, Arachis hypogaea L., conhecidos como Roxo, Tatu Branco, Tatu Vermelho, Tatuí Vermelho e Tatuí (sementes com película branca), fornecidos por fazendas situadas nas regiões dos municípios de Marília, Presidente Prudente e São Manuel. Para tal análise, foi utilizada a técnica de eletroforese horizontal em gel de poliacrilamida, para os sistemas da leucil-aminopeptidase (LAP), aspartato aminotransferase (ATT) e peroxidase (PER). No sistema da leucil-aminopeptidase, foram observadas três bandas enzimáticas, denominadas LAP-A, LAP-B e LAP-C. Os padrões de bandas obtidos para o sistema da aspartato-aminotransferase mostraram a existência de três bandas anódicas, AAT-A, AAT-B e AAT-C. No sistema da peroxidase (PER), foram observadas quinze bandas, sendo oito anódicas (PER-A a PER-H) e sete catódicas (PER-I a PER-P). Os sistemas enzimáticos da peroxidase e leucil-aminopeptidase não foram discriminativos para as amostras analisadas dos diferentes cultivares obtidos nas diversas regiões. O sistema da aspartato-aminotransferase apresentou um padrão composto pelas bandas AAT-B e AAT-C, que se mostrou característico e discriminativo para as amostras do cultivar Tatu Branco, procedente de Presidente Prudente, e do 'Tatuí Vermelho', proveniente de Presidente Prudente e São Manuel.

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The present investigation was undertaken to identify and characterize trophozoite proteases of five axenic strains of Giardia duodenalis isolated in Brazil and the reference strain Portland 1 isolated in the United States. Trophozoite cell lysates of each strain were analysed for the pattern of proteins and for proteolytic activity. Samples were tested in SDS-polyacrylamide gel electrophoresis for the protein profiles, and the detection of proteases in cell lysates was performed using substrate gel electrophoresis [gelatin, collagen, bovine serum albumin (BSA) and haemoglobin] and azocasein assays. Indeed, synthetic inhibitors were included in the assays to characterize the protease classes. Differences on the hydrolysis patterns of protein substrates were observed in relation to the substrate composition as much as the Giardia trophozoite strain. The substrate-containing gels revealed hydrolysis bands with molecular masses ranging from > 97 to 20-15 kDa, and most zones were common to the five strains. However, some pronounced differences could be detected in the BTU-11 pattern. Azocasein was also degraded; however, depending on the lysate assayed, the degree of substrate degradation was variable. It was observed that inhibitory effects are substrate-dependent since the activity was predominantly due to cysteine proteases against gelatin, collagen, BSA and azocasein substrates and due to serine against haemoglobin. The presence of aspartic protease and aminopeptidase activity in the lysates was also indicated.

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The adult female Culex quinquefasciatus midgut comprises a narrow anterior and a dilated posterior region, with epithelia composed of a monolayer of adjacent epithelial cells joined at the apical portion by septate junctions. Densely packed apical microvilli and an intricate basal labyrinth characterise each cell pole. Our morphological studies suggest that, during blood digestion, the anterior midgut region also participates in an initial absorptive stage which is probably related to the intake of water, salts and other small molecules. This activity peaked by 6 h after bloodmeal feeding (ABF) and ended approximately 18 h ABF, when the peritrophic membrane was already formed. After this time, absorption only occurred in the posterior region, with morphologic and biochemical evidence of high synthetic activity related to the secretion of proteases. Chymotrypsin, elastase, aminopeptidase, and trypsin reached their maximum activity at around 36 h ABF. Digestion products were apparently absorbed and transported to the basal labyrinth, from where they should be released to the hemolymph. At 72 h ABF, proteolysis had already ended and protein levels had returned to those observed before blood meal. The epithelium of the posterior region, however, did not return to its initial morphology, appearing quite disorganised. Additionally, from 48 h ABF onwards some epithelial cells showed morphological signals of apoptosis. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

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

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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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)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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