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

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

Author Silva, Mariana Cristina Cabral Autor UNIFESP Google Scholar
Santana, Lucimeire Aparecida de Autor UNIFESP Google Scholar
Mentele, Reinhard Google Scholar
Ferreira, Rodrigo da Silva Autor UNIFESP Google Scholar
Miranda, Antonio Autor UNIFESP Google Scholar
Silva-Lucca, Rosemeire Aparecida Google Scholar
Sampaio, Misako Uemura Autor UNIFESP Google Scholar
Correia, Maria Tereza dos Santos Google Scholar
Oliva, Maria Luiza Vilela Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Estadual Oeste Parana
Universidade Federal de Pernambuco (UFPE)
Inst Clin Neuroimmunol LMU
Max Planck Inst Biochem
Abstract 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.
Keywords Bauhinia forficata
Coagulation time
Plant lectin
Platelet aggregation
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Grant number FAPESP: 07/58929-4
FAPESP: 09/53766-5
Date 2012-07-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 47, n. 7, p. 1049-1059, 2012.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 1049-1059
Origin http://dx.doi.org/10.1016/j.procbio.2012.03.008
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000309333600005
URI http://repositorio.unifesp.br/handle/11600/35013

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