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|Title:||Purification, characterization, and preliminary x-ray diffraction analysis of a lactose-specific lectin from Cymbosema roseum seeds|
|Authors:||Rocha, Bruno Anderson Matias da|
Moreno, Frederico Bruno Mendes Batista
Souza, Emmanuel Prata
Marinho, Emmanuel S.
Benevides, R. G.
Rustiguel, Joane Kathelen Rodrigues
Souza, Luis A.G.
Nagano, C. S.
Sampaio, Alexandre Holanda
Azevedo, Walter Filgueira de
Cavada, B. S.
X Ray Diffraction Analysis
Isolation And Purification
Tandem Mass Spectrometry
Unit Cell Parameters
Amino Acid Sequence
Protein Carbohydrate Interaction
|metadata.dc.publisher.journal:||Applied Biochemistry and Biotechnology|
|metadata.dc.relation.ispartof:||Volume 152, Número 3, Pags. 383-393|
|Abstract:||The unique carbohydrate-binding property of lectins makes them invaluable tools in biomedical research. Here, we report the purification, partial primary structure, carbohydrate affinity characterization, crystallization, and preliminary X-ray diffraction analysis of a lactose-specific lectin from Cymbosema roseum seeds (CRLII). Isolation and purification of CRLII was performed by a single step using a Sepharose-4B-lactose affinity chromatography column. The carbohydrate affinity characterization was carried using assays for hemagglutination activity and inhibition. CRLII showed hemagglutinating activity toward rabbit erythrocytes. O-glycoproteins from mucine mucopolysaccharides showed the most potent inhibition capacity at a minimum concentration of 1.2 μg mL-1. Protein sequencing by mass spectrometry was obtained by the digestion of CRLII with trypsin, Glu-C, and AspN. CRLII partial protein sequence exhibits 46% similarity with the ConA-like α chain precursor. Suitable protein crystals were obtained with the hanging-drop vapor-diffusion method with 8% ethylene glycol, 0.1 M Tris-HCl pH 8.5, and 11% PEG 8,000. The monoclinic crystals belong to space group P21 with unit cell parameters a∈=∈49.4, b∈=∈89.6, and c∈=∈100.8 Å. © 2008 Humana Press.|
|Appears in Collections:||Artigos|
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