Purification, Partial Characterization, and CNBr-Sepharose Immobilization of a Vasorelaxant Glucose/Mannose Lectin from Canavalia virosa Seeds

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-me...

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 2014-04, Vol.172 (7), p.3342-3353
Main Authors: Osterne, Vinicius J. S, Santiago, Mayara Q, Pinto-Junior, Vanir R, Cajazeiras, João B, Correia, Jorge L. A, Leitão, Cintia C. F, Carneiro, Rômulo F, Pereira-Junior, Francisco N, Vasconcelos, Mayron A, Rocha, Bruno A. M, Assreuy, Ana Maria S, Bringel, Pedro Henrique S. F, Nagano, Celso S, Nascimento, Kyria S, Cavada, Benildo S
Format: Article
Language:English
Subjects:
affinity chromatography
Animals
aorta
Aorta - drug effects
Aorta - physiopathology
Artemia
Artemia - drug effects
Biochemistry
Biological and medical sciences
Biotechnology
Canavalia - chemistry
Canavalia cathartica
Chemistry
Chemistry and Materials Science
Chromatography
Chromatography, Affinity
EDTA (chelating agent)
erythrocytes
fetuins
Fundamental and applied biological sciences. Psychology
General aspects
glucose
Glucose - metabolism
glycoproteins
Hemagglutination
Immobilization techniques
lectins
Legumes
mannose
Mannose - metabolism
mass spectrometry
Methods. Procedures. Technologies
nauplii
ovalbumin
Plant Lectins - chemistry
Plant Lectins - isolation & purification
Plant Lectins - metabolism
polyacrylamide gel electrophoresis
Protein Stability
Proteins
Rabbits
Rats
Rats, Wistar
Seeds
Seeds - chemistry
toxicity
Vasodilator Agents - chemistry
Vasodilator Agents - isolation & purification
Vasodilator Agents - metabolism
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Description
Summary: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.
ISSN:0273-2289
1559-0291
DOI:10.1007/s12010-014-0751-3