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Purification, Partial Characterization and Immobilization of a Mannose-Specific Lectin from Seeds of Dioclea lasiophylla Mart.
Laboratory of Biologically Active Molecules (Biomol-Lab), Department of Biochemistry and Molecular Biology, Federal University of Ceará, Av. Humberto Monte s/n, Bloco 907, Lab. 1075, Campus do Pici, Fortaleza-CE 60440-970, Brazil
Integrated Laboratory of Biomolecules (LIBS), Federal University of Ceará, Department of Pathology and Legal Medicine, Faculty of Medicine, Fortaleza, CE 60430-160, Brazil
Laboratory of Mass Spectrometry Applied to Proteins (LEMAP), Federal University of Ceará, Av. Humberto Monte s/n, Bloco 825, Campus do Pici, Fortaleza-CE 60440-970, Brazil
* Author to whom correspondence should be addressed.
Received: 31 July 2013; in revised form: 29 August 2013 / Accepted: 30 August 2013 / Published: 4 September 2013
Abstract: Lectin from the seeds of Dioclea lasiophylla (DlyL) was purified in a single step by affinity chromatography on a Sephadex® G-50 column. DlyL strongly agglutinated rabbit erythrocytes and was inhibited by monosaccharides (D-mannose and α-methyl-D-mannoside) and glycoproteins (ovalbumin and fetuin). Similar to other Diocleinae lectins, DlyL has three chains, α, β and γ, with mass of 25,569 ± 2, 12,998 ± 1 and 12,588 ± 1 Da, respectively, and has no disulfide bonds. The hemagglutinating activity of DlyL was optimal in pH 8.0, stable at a temperature of 70 °C and decreased in EDTA solution, indicating that lectin activity is dependent on divalent metals. DlyL exhibited low toxicity on Artemia sp. nauplii, but this effect was dependent on the concentration of lectin in solution. DlyL immobilized on cyanogen bromide-activated Sepharose® 4B bound 0.917 mg of ovalbumin per cycle, showing the ability to become a tool for glycoproteomics studies.
Keywords: lectin; Dioclea lasiophylla; Diocleinae; toxicity; immobilization
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Pinto-Júnior, V.R.; de Santiago, M.Q.; Osterne, V.J.S.; Correia, J.L.A.; Pereira-Júnior, F.N.; Cajazeiras, J.B.; de Vasconcelos, M.A.; Teixeira, E.H.; do Nascimento, A.S.F.; Miguel, T.B.A.R.; Miguel, E.C.; Sampaio, A.H.; do Nascimento, K.S.; Nagano, C.S.; Cavada, B.S. Purification, Partial Characterization and Immobilization of a Mannose-Specific Lectin from Seeds of Dioclea lasiophylla Mart.. Molecules 2013, 18, 10857-10869.
Pinto-Júnior VR, de Santiago MQ, Osterne VJS, Correia JLA, Pereira-Júnior FN, Cajazeiras JB, de Vasconcelos MA, Teixeira EH, do Nascimento ASF, Miguel TBAR, Miguel EC, Sampaio AH, do Nascimento KS, Nagano CS, Cavada BS. Purification, Partial Characterization and Immobilization of a Mannose-Specific Lectin from Seeds of Dioclea lasiophylla Mart.. Molecules. 2013; 18(9):10857-10869.
Pinto-Júnior, Vanir R.; de Santiago, Mayara Q.; Osterne, Vinícius J.S.; Correia, Jorge L.A.; Pereira-Júnior, Francisco N.; Cajazeiras, João B.; de Vasconcelos, Mayron A.; Teixeira, Edson H.; do Nascimento, Antônia S.F.; Miguel, Thaiz B.A.R.; Miguel, Emilio C.; Sampaio, Alexandre H.; do Nascimento, Kyria S.; Nagano, Celso S.; Cavada, Benildo S. 2013. "Purification, Partial Characterization and Immobilization of a Mannose-Specific Lectin from Seeds of Dioclea lasiophylla Mart.." Molecules 18, no. 9: 10857-10869.