Title: Purification and Partial Characterization of a New Pro-Inflammatory Lectin from Bauhinia bauhinioides Mart (Caesalpinoideae) Seeds
VOLUME: 18 ISSUE: 4
Author(s):Helton C. Silva, Alfa U. Bari, Francisco N. Pereira-Junior, Rafael C. Simoes, Ito L. Barroso-Neto, Camila B. Nobre, Maria G. Pereira, Kyria S. Nascimento, Bruno Anderson M. Rocha, Plinio Delatorre, Celso S. Nagano, Ana Maria S. Assreuy and Benildo S. Cavada
Affiliation:Departamento de Bioquimica e Biologia Molecular, Laboratorio de Moleculas Biologicamente Ativas, Universidade Federal do Ceara, Campus do Pici, s/n Bloco 907, Caixa Postal 6043, CEP 60.455-970, Fortaleza-CE, Brasil.
Keywords:Lectin, bauhinia bauhinioides, caesalpinoideae, pro-inflammatory activity, galactose-specific lectin, proteolytic enzymes, L-NAME, Lectin Purification, Molecular Mass Determination, Electrospray ionization, mass spectrometry (ESI-MS), bovine serum albumin (BSA), Rat Paw Edema Model, -methyl-D-galactopyranoside, HPLC, BBLLectin, bauhinia bauhinioides, caesalpinoideae, pro-inflammatory activity, galactose-specific lectin, proteolytic enzymes, L-NAME, Lectin Purification, Molecular Mass Determination, Electrospray ionization, mass spectrometry (ESI-MS), bovine serum albumin (BSA), Rat Paw Edema Model, -methyl-D-galactopyranoside, HPLC, BBL
Abstract: A new galactose-specific lectin, named BBL, was purified from seeds of Bauhinia bauhinioides by precipitation with ammonium sulfate, followed by two steps of ion exchange chromatography. BBL haemagglutinated rabbit erythrocytes (native and treated with proteolytic enzymes) showing stability even after exposure to 60 °C for one hour. The lectin haemagglutinating activity was optimum between pH 8.0 and 9.0 and inhibited after incubation with D-galactose and its derivatives, especially α-methyl-D-galactopyranoside. The pure protein possessed a molecular mass of 31 kDa by SDS – PAGE and 28.310 Da by mass spectrometry. The lectin pro-inflammatory activity was also evaluated. The s.c. injection of BBL into rats induced a dose-dependent paw edema, an effect that occurred via carbohydrate site interaction and was significantly reduced by L-NAME, suggesting an important participation of nitric oxide in the late phase of the edema. These findings indicate that BBL can be used as a tool to better understand the mechanisms involved in inflammatory responses.
