Isolation and characterization of a lectin from the seeds of Dioclea grandiflora (Mart.)

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Abstract
By a combination of solubility fractionation, affinity and molecular-sieve chromatography, a lectin preparation containing several closely related lectin components of different isoelectric point was isolated from the seeds of Dioclea grandiflora Mart. The lectins showed a carbohydrate specificty for D-mannose (D-glucose)-binding and had a requirement for the presence of Ca2+ and Mn2+. The results of preliminary characterization studies showed that the D. grandiflora lectins had similar properties to those of concanavalin A, the lectin from the seeds of Canavalia ensiformis, a plant also belonging to the tribe Diocleae. Thus the D. grandiflora lectins contained no covalently bound carbohydrate and had an amino-acid composition characterized by a low content of methionine and the virtual absence of cysteine. Above pH 4.8 they had molecular weight of about 100,000, while below pH 3.1 they were dissociated to half-molecules. Between these two pH values there was a fast association-dissociation equilibrium for the two species. In dissociating solvents, three subunits were obtained of the approximate size of 25–26,000, 13–14,000 and 8–9,000. The lectins from C. grandiflora similar to concanavalin A were more distantly related to the lectins obtained from the members of the tribe Vicieae although these were also specific for D-mannose (D-glucose)-binding.