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A novel vasorelaxant lectin purified from seeds of Clathrotropis nitida: Partial characterization and immobilization in chitosan beads
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Abstract:
A novel lectin from seeds of Clathrotropis nitida (CNA) was purified and characterized. CNA is a glycoprotein containing approximately 3.3% carbohydrates in its structure. CNA promoted intense agglutination of rabbit erythrocytes, which was inhibited by galactosides and porcine stomach mucin (PSM). The lectin maintained its hemagglutinating activity after incubation in a wide range of temperatures (30-60 °C) and pH (6.0-7.0), and its binding activity was dependent on divalent cations (Ca+2 and Mg+2). SDS-PAGE showed an electrophoretic profile consisting of a single band of 28 kDa, as confirmed by electrospray ionization mass spectrometry, which indicated an average molecular mass of 27,406 ± 2 Da and the possible presence of isoforms and glycoforms. In addition, CNA exhibited no toxicity to Artemia sp. nauplii and elicited reversible and dose-dependent vasorelaxation in precontracted aortic rings. CNA was successfully immobilized on chitosan beads and was able to capture PSM in solution. This study demonstrated that CNA is a lectin that has potential as a biotechnological tool in glycomics and glycoproteomics applications. © 2015 Elsevier Inc.
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Calcium, Carbohydrate, Chitosan, Galactoside, Lectin, Magnesium, Stomach Mucin, Immobilized Protein, Plant Lectin, Vasodilator Agent, Animals Cell, Animals Experiment, Animals Model, Binding Affinity, Carbohydrate Analysis, Clathrotropis Nitida, Controlled Study, Dose Response, Electrospray Mass Spectrometry, Glycobiology, Male, Medicinal Plant, Molecular Weight, Nonhuman, Physical Chemistry, Seed Plant, Electrophoresis, Polyacrylamide Gel, Priority Journal, Protein Analysis, Protein Immobilization, Rat, Temperature, Vascular Ring, Vasodilatation, Amino Acid Sequence, Animals, Artemia, Chemistry, Drug Effects, Fabaceae, Genetics, Hemagglutination, Human, In Vitro Study, Isolation And Purification, Molecular Genetics, Physiology, Rabbit, Sequence Homology, Aorta, Thoracic, Wistar Rat, Amino Acid Sequence, Animal, Aorta, Thoracic, Artemia, Chitosan, Fabaceae, Hemagglutination, Humans, Immobilized Proteins, In Vitro Techniques, Male, Molecular Sequence Data, Molecular Weight, Plant Lectins, Plants, Medicinal, Rabbits, Rats, Rats, Wistar, Seeds, Sequence Homology, Amino Acid, Vasodilator Agents
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