Purification, Partial Characterization, and CNBr-Sepharose Immobilization of a Vasorelaxant Glucose/Mannose Lectin from Canavalia virosa Seeds
| Autor: | Vanir Reis Pinto-Junior, Francisco N. Pereira-Junior, Celso Shiniti Nagano, Ana Maria Sampaio Assreuy, Mayara Queiroz Santiago, Vinicius Jose Da Silva Osterne, Cintia C. F. Leitão, Jorge Luis Almeida Correia, Benildo Sousa Cavada, Mayron Alves de Vasconcelos, Rômulo Farias Carneiro, Kyria S. Nascimento, Bruno A.M. Rocha, Pedro Henrique de Souza Ferreira Bringel, João Batista Cajazeiras |
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| Rok vydání: | 2014 |
| Předmět: |
Vasodilator Agents
Mannose Bioengineering Applied Microbiology and Biotechnology Biochemistry Chromatography Affinity Sepharose chemistry.chemical_compound Affinity chromatography Animals Monosaccharide Rats Wistar Molecular Biology Aorta chemistry.chemical_classification Chromatography biology Protein Stability Hemagglutination Lectin General Medicine Canavalia biology.organism_classification Fetuin Rats Ovalbumin Glucose chemistry Seeds biology.protein Rabbits Artemia Plant Lectins Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 172:3342-3353 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | A novel mannose/glucose-binding lectin from Canavalia virosa (designated as ConV) has been purified from seeds of C. virosa by affinity chromatography on a mannose-Sepharose 4B column. ConV strongly agglutinates rabbit erythrocytes and was inhibited by monosaccharides (D-mannose, D-glucose, and α-methyl-D-mannoside) and glycoproteins (ovalbumin and fetuin). SDS-PAGE revealed three bands corresponding to three subunits (α, β, and γ) confirmed by ESI mass spectrometry with exact mass of 25,480 ± 2 Da, 12,864 ± 1 Da, and 12,633 ± 1 Da, respectively. The purified lectin was more stable in pH ranging from 7.0 to 9.0, supported up to 80 ºC without any loss in activity and unaffected by EDTA. ConV showed no toxicity against Artemia sp. nauplii and relaxed endothelized rat aorta, with the participation of the lectin domain. In our tests, the lectin immobilized on CNBr-Sepharose was capable of binding 0.8 mg of ovalbumin per chromatography, allowing the use of ConV as a tool for capture and purification of glycoproteins. |
| Databáze: | OpenAIRE |
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