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

• DOI: 10.1016/j.procbio.2012.03.008
• Publication Date: 2012-04-04T03:46:32Z
• Authors: Mariana C.C. Silva; Lucimeire A. Santana; Reinhard Mentele; Rodrigo S. Ferreira; Antônio de Miranda; Rosemeire A. Silv...; Misako U. Sampaio; Maria T.S. Correia; Maria L.V. Oliva
• Keywords: Bauhinia; Sephadex; Molecular mass

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 Superdex 75 size exclusion chromatography. The molecular homogeneity purity of BfL were assessed reversed-phase HPLC. appeared as a single band approximately 27.0 kDa on SDS-PAGE under non-reducing reducing conditions, its weight determined to be 27,850 Da LC/ESI-MS. is glycoprotein with carbohydrate content 6.24% the phenol–sulfuric acid method. Fetuin, asialofetuin, thyroglobulin azocasein inhibited hemagglutinating activity whereas saccharides did not. stable at 100 °C for 30 min, pH-dependent, highest pH 6.0, metal-independent. primary structure shows similarity other lectins genus Bauhinia. Deconvolution circular dichroism (CD) spectrum indicated presence α-helix β structures. increases coagulation time, but this effect not related human plasma kallikrein or factor Xa inhibition. also inhibits ADP- epinephrine-induced platelet aggregation in dose-dependent manner only currently described lectin that exhibits anticoagulant antiplatelet aggregating properties.