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Molecules ; 16(6): 5087-103, 2011 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-21694673

RESUMEN

DwL, a lectin extracted from the seeds of Dioclea wilsonii, is a metalloprotein with strong agglutinating activity against rabbit and ABO erythrocytes, inhibited by glucose and mannose. DwL was purified by affinity chromatography on a Sephadex G-50 column and ion exchange chromatography on a HiTrap SP XL column. SDS-PAGE revealed three electrophoretic bands corresponding to the α (25,634 ± 2 Da), ß (12,873 ± 2 Da) and γ (12,779 ± 2 Da) chains. Protein sequencing was done by Tandem Mass Spectrometry. The primary sequence featured 237 amino acids and was highly homologous to other reported Diocleinae lectins. A complete X-ray dataset was collected at 2.0 Å for X-Man-complexed DWL crystals produced by the vapor diffusion method. The crystals were orthorhombic and belonged to the space group I222, with the unit-cell parameters a = 59.6, b = 67.9 and c = 109.0 Å. DWL differed in potency from other ConA-like lectins and was found to induce neutrophil migration in rats, making it particularly useful in structural/functional studies of this class of proteins.


Asunto(s)
Dioclea/química , Mediadores de Inflamación/química , Lectinas de Plantas/química , Semillas/química , Secuencia de Aminoácidos , Animales , Movimiento Celular/efectos de los fármacos , Secuencia Conservada , Cristalización , Eritrocitos/efectos de los fármacos , Humanos , Mediadores de Inflamación/aislamiento & purificación , Mediadores de Inflamación/farmacología , Datos de Secuencia Molecular , Neutrófilos/efectos de los fármacos , Lectinas de Plantas/aislamiento & purificación , Lectinas de Plantas/farmacología , Estabilidad Proteica , Conejos , Ratas , Ratas Wistar , Alineación de Secuencia
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