Purification, primary structure and potential functions of a novel lectin from Bauhinia forficata seeds

• Autor(a) principal: Silva, Mariana Cristina Cabral [UNIFESP]
• Data de Publicação: 2012
• Outros Autores: Santana, Lucimeire Aparecida de [UNIFESP], Mentele, Reinhard, Ferreira, Rodrigo da Silva [UNIFESP], Miranda, Antonio [UNIFESP], Silva-Lucca, Rosemeire Aparecida, Sampaio, Misako Uemura [UNIFESP], Correia, Maria Tereza dos Santos, Oliva, Maria Luiza Vilela [UNIFESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://dx.doi.org/10.1016/j.procbio.2012.03.008; http://repositorio.unifesp.br/handle/11600/35013
• topic: Bauhinia forficata; Caesalpinoideae; Coagulation time; Plant lectin; Platelet aggregation; Seeds

A new lectin. Bfl. was purified from Bauhinia forficata seeds by ammonium sulfate fractionation. DEAE-Sephadex ion exchange chromatography, Sepharose-4B and chitin affinity chromatographies and Superdex 75 size exclusion chromatography. the molecular homogeneity and purity of BfL were assessed by reversed-phase H PLC. BfL appeared as a single band of approximately 27.0 kDa on SDS-PAGE under non-reducing and reducing conditions, and its molecular weight was determined to be 27,850 Da by LC/ESI-MS. Bit is a glycoprotein with a carbohydrate content of 6.24% determined by the phenol-sulfuric acid method. Fetuin, asialofetuin, thyroglobulin and azocasein inhibited the hemagglutinating activity of BfL, whereas saccharides did not. Bfl hemagglutinating activity was stable at 100 degrees C for 30 min, pH-dependent, with the highest activity at pH 6.0, and metal-independent. the primary structure of BfL shows similarity with other lectins from the genus Bauhinia. Deconvolution of the BfL circular dichroism (CD) spectrum indicated the presence of alpha-helix and beta structures. BfL increases coagulation time, but this effect is not related to human plasma kallikrein or human factor Xa inhibition. Bfl also inhibits ADP- and epinephrine-induced platelet aggregation in a dose-dependent manner and is the only currently described lectin from Bauhinia that exhibits anticoagulant and antiplatelet aggregating properties. (c) 2012 Elsevier B.V. All rights reserved.