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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Structural and Biological Characterization of a Seed Lectin from Centrolobium tomentosum Guill ex. Benth / CaracterizaÃÃo estrutural e biolÃgica de uma lectina de sementes de centrolobium tomentosum guill. ex benth

Alysson Chaves Almeida 18 February 2016 (has links)
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A glycosylated lectin (CTL) with specificity for mannose and glucose has been detected and purified from seeds of Centrolobium tomentosum, a legume plant from the Dalbergieae tribe. CTL was isolated by mannose-Sepharose affinity chromatography. The primary structure was determined by tandem mass spectrometry and consists of 245 amino acids and one N-glycosylation site, possessing high similarity with the lectin Platypodium elegans (PELa) and Pterocarpus angolensis (PAL) derived from the same tribe. Two crystal structures of CTL, with monoclinic and tetragonal forms, both complexed with methyl dimanosÃdeo has been solved at 2.25 and 1.9 Ã, respectively, with high similarity. The lectin adopts a typical canonical dimeric organization of legume lectins. The carbohydrate recognition domain (CRD), metal binding site, and glycosylation site have been characterized and the structural basis for interaction with carbohydrate been elucidated. CTL showed acute inflammatory effect in a paw edema model. The protein structure was subjected to ligand screening (dimannosides and trimannoside) by molecular docking, revealing a higher affinity for trimannosides and their interactions were compared with similar lectins, which have the same binding specificity. This is the first report of a crystal structure of a native mannose lectin / specific glucose Dalbergieae tribe with pro-inflammatory activity / Uma lectina glicosilada (CTL) com especificidade a manose e glucose foi detectada e purificada a partir de sementes de Centrolobium Tomentosum, uma leguminosa pertencente à tribo Dalbergieae. CTL foi isolada por cromatografia de afinidade de Sepharose-Manose. A estrutura primÃria foi determinada por espectrometria de massas e consiste em 245 aminoÃcidos e um sitio de ÂNÂ-glicosilaÃÃo, demonstrando similaridade com a lectina de Platypodium elegans (PELa), Pterocarpus angolensis (PAL), dentre outras, oriundas da mesma tribo. Duas estruturas cristalinas de CTL, de formas monoclÃnica e tetragonal, ambas complexadas com metil-dimanosÃdeo, foram resolvidas a 2,25 e 1,9 Ã, respectivamente, apresentando alta similaridade entre si. A lectina mostrou adotar uma organizaÃÃo dimÃrica canÃnica tÃpica de lectinas de leguminosas. O domÃnio de reconhecimento de carboidratos (CRD), local de ligaÃÃo do metal e local de glicosilaÃÃo foram caracterizados e a base estrutural para a interaÃÃo com carboidratos foi elucidado. CTL mostrou efeito inflamatÃrio agudo em um modelo de edema de pata. A estrutura da proteÃna foi submetida a uma anÃlise de interaÃÃes com dimanosÃdeos e trimanosÃdeos por Docking Molecular, revelando sua maior afinidade por trimanosÃdeos e suas interaÃÃes foram comparadas com lectinas similares que possuam a mesma especificidade de ligaÃÃo. Esse à o primeiro relato de estrutura cristalina de uma lectina nativa manose/glucose especÃfica da tribo Dalbergieae com atividade prÃ-inflamatÃria

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