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Applied Biochemistry and Biotechnology

, Volume 172, Issue 7, pp 3342–3353 | Cite as

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

  • Vinicius J. S. Osterne
  • Mayara Q. Santiago
  • Vanir R. Pinto-Junior
  • João B. Cajazeiras
  • Jorge L. A. Correia
  • Cintia C. F. Leitão
  • Rômulo F. Carneiro
  • Francisco N. Pereira-Junior
  • Mayron A. Vasconcelos
  • Bruno A. M. Rocha
  • Ana Maria S. Assreuy
  • Pedro Henrique S. F. Bringel
  • Celso S. Nagano
  • Kyria S. Nascimento
  • Benildo S. CavadaEmail author
Article

Abstract

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.

Keywords

Canavalia virosa Lectin Diocleinae Purification Immobilization Vascular smooth muscle Relaxant activity 

Notes

Acknowledgement

Research supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), and Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP); David Harding helped with English editing of the manuscript. Assreuy AMS, Rocha BAM, Nascimento KS, and Cavada BS are senior investigators of CNPq/Brazil.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Vinicius J. S. Osterne
    • 1
  • Mayara Q. Santiago
    • 1
  • Vanir R. Pinto-Junior
    • 1
  • João B. Cajazeiras
    • 1
  • Jorge L. A. Correia
    • 1
  • Cintia C. F. Leitão
    • 1
  • Rômulo F. Carneiro
    • 1
  • Francisco N. Pereira-Junior
    • 1
  • Mayron A. Vasconcelos
    • 1
  • Bruno A. M. Rocha
    • 1
  • Ana Maria S. Assreuy
    • 2
  • Pedro Henrique S. F. Bringel
    • 2
  • Celso S. Nagano
    • 3
  • Kyria S. Nascimento
    • 1
  • Benildo S. Cavada
    • 1
    • 4
    Email author
  1. 1.BioMol-Lab - Laboratório de Moléculas Biologicamente Ativas, Departamento de Bioquímica e Biologia MolecularUniversidade Federal do CearáFortalezaBrazil
  2. 2.LAFFIN – Laboratório de Fisio-Farmacologia da Inflamação, Instituto Superior de Ciências BiomédicasUniversidade Estadual do CearáFortalezaBrazil
  3. 3.LEMAP - Laboratório de Espectrometria de Massas Aplicado à Proteínas, Departamento de Engenharia de PescaUniversidade Federal do CearáFortalezaBrazil
  4. 4.FortalezaBrazil

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