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

Autor(es)

Silva, Mariana C. C. ^[1] ; Santana, Lucimeire A. ^[1] ; Mentele, Reinhard ^{[2, 3]} ; Ferreira, Rodrigo S. ^[1] ; de Miranda, Antonio ^[4] ; Silva-Lucca, Rosemeire A. ^[5] ; Sampaio, Misako U. ^[1] ; Correia, Maria T. S. ^[6] ; Oliva, Maria L. V. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Bioquim, BR-04044020 Sao Paulo - Brazil

^[2] Inst Clin Neuroimmunol LMU, Munich - Germany

^[3] Max Planck Inst Biochem, Dept Prot Analyt, D-82152 Martinsried - Germany

^[4] Univ Fed Sao Paulo, Dept Biofis, BR-04044020 Sao Paulo - Brazil

^[5] Univ Estadual Oeste Parana, Ctr Engn & Ciencias Exatas, BR-85903000 Toledo, PR - Brazil

^[6] Univ Fed Pernambuco, Dept Bioquim, BR-50670901 Recife, PE - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Process Biochemistry; v. 47, n. 7, p. 1049-1059, JUL 2012.
• Citações Web of Science: 20

Resumo

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 Ltd. All rights reserved. (AU)

• Processo FAPESP: 07/58929-4 - Purificacao, caracterizacao bioquimica e fisiologica de lectinas de plantas do genero bauhinia e sua acao sobre linhagens celulares tumorigenicas.
• Beneficiário: Mariana Cristina Cabral Silva
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 09/53766-5 - Proteínas de origem vegetal com seletividade para inibição de enzimas de mamíferos e seu papel como agente anti-inflamatório, antitrombótico, antidiabético e antitumoral
• Beneficiário: Maria Luiza Vilela Oliva
• Modalidade de apoio: Auxílio à Pesquisa - Temático