Purification and primary structure determination of a galactose-specific lectin from Vatairea guianensis Aublet seeds that exhibits vasorelaxant effect |
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Authors: | Helton C Silva Celso S Nagano Luis AG Souza Kyria S Nascimento Renato Isídro Plínio Delatorre Bruno Anderson M Rocha Alexandre H Sampaio Ana Maria S Assreuy Alana F Pires Luis Eduardo A Damasceno Gabriela FO Marques-Domingos Benildo S Cavada |
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Institution: | 1. BioMol-Lab, Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, P.O. Box 6043, 60440-970 Fortaleza, Ceará, Brazil;2. Laboratório de Espectrometria de Massa Aplicado a Proteínas (LEMAP/Biomol-Lab), Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Av. Humberto Monte s/n, Bloco 825, Campus do Pici, P.O. Box 6043, 60440-970 Fortaleza, Ceará, Brazil;3. Instituto Nacional de Pesquisas da Amazônia-INPA, Manaus, Amazonas, Brazil;4. Departamento de Biologia Molecular, Universidade Federal da Paraíba, João Pessoa, Brazil;5. Instituto Superior de Ciências Biomédicas, Universidade Estadual do Ceará, Fortaleza, Brazil |
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Abstract: | Vatairea guianensis seeds, a typical plant from the Brazilian Amazon region that belongs to the Dalbergieae tribe, possess a lectin that was isolated by precipitation with solid ammonium sulfate followed by guar gum affinity chromatography. This lectin was named VGL. The V. guianensis lectin strongly agglutinated rabbit erythrocytes and was inhibited by d-galactose and d-galactose-derived sugars, especially N-acetyl-d-galactosamine. VGL has been shown to be a stable protein, maintaining its hemagglutinating activity after incubation at a wide range of temperature and pH values and after incubation with ethylenediamine tetraacetic acid (EDTA). In a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, the purified VGL exhibited an electrophoretic profile consisting of a major 30–32 kDa double band, which is termed the alpha-chain, and two minor components of 18 and 15 kDa, which are referred to as the beta- and gamma-chains, respectively. An analysis using electrospray ionization mass spectrometry also indicated that purified VGL contains a mixture of chains with molecular weights of 28,437 ± 2, 14,952 ± 2 and 12,332 ± 2. The complete amino acid sequence of VGL, as determined using tandem mass spectrometry, consists of 239 amino acid residues. VGL is a glycoprotein exhibiting high similarity in primary structure to other lectins from evolutionarily related plants, such as Vatairea macrocarpa lectin and lectins belonging to the Sophoreae tribe. VGL exhibits vasorelaxant activity in contracted rat aortas, an effect that is strictly dependent on the endothelium and involves nitric oxide and the lectin domain. |
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