Lectinas solubles beta-galactosídicas / Beta-galactoside solubles lectins

Sobre la base de estudios estructurales y funcionales, las lectinas animales se han clasificado en dos tipos: el Tipo C, caracterizado por su dependencia de los iones de calcio y el Tipo S que no es calciodependiente, sino tioldependiente. Entre estas últimas, se ha estudiado ampliamente el grupo de lectinas S-Lac, que son extraídas con buffers salinos con lactosa, en presencia de tioles. Contituyen una familia de proteínas realcionadas estructuralmente, que contienen una serie de aminoácidos conservados. Se unen específicamente a glicoconjugados complementarios y su biosíntesis y localización son reguladas por el desarrolo. Su rol puede relacionarse con diversas actividades biológicas que poderían variar según el órgano.

Identification of endogenous soluble glycoprotein receptors of bovine brain grey matter 14 kDa beta-galactoside-binding lectin.

The 14 kDa beta-galactoside-binding lectin from bovine brain grey matter (BBL) covalently attached to caproic acid-Sepharose by the N-hydroxy succinimide procedure was used to isolate endogenous glycoprotein receptors of this lectin. BBL-Sepharose could sugar-specifically retain several endogenous soluble glycoproteins with subunit molecular mass (in kDA) 44, 51, 60, 123 and 186. BBL, conjugated with horse radish peroxidase, could sugar-specifically recognize several glycoprotein subunits with molecular mass (in kDA) 58, 87, and 117 and 186 on Western blots. The only protein from an extract of bovine brain grey matter, that retained on Sepharose-immobilized endogenous N-linked glycoproteins and subsequently eluted with beta-galactosides was BBL as confirmed by electrophoresis and agglutination inhibition measurement. N-linked glycoproteins from bovine heart and even from human placenta were also efficient receptors of BBL. These results suggest that 14 kDa beta-galactoside-binding lectin is the major protein, if not the only one, that sugar-specifically interacts with endogenous soluble glycoproteins in bovine brain grey matter.

Sugar-specific interaction of bovine brain beta-galactoside-binding lectin with endogenous gangliosides.

Sugar-specific binding of the 14 kDa beta-galactoside-binding lectin from bovine brain grey matter to mixed endogenous gangliosides was demonstrated by affinity electrophoresis and hemagglutination inhibition. Gangliosides prepared by Folch extraction, base treatment and silica gel chromatography, when incorporated in native or desialated form in polyacrylamide gel above their critical micellar concentration, arrested the mobility of the lectin during electrophoresis at pH 8.2. This effect was sugar-specific since it was reversed if lactose, but not sucrose, was present in the gel. Also, retention of the brain lectin by ganglioside and its reversal by lactose were concentration-dependent. In presence of bovine serum albumin, at pH 7.4 native and desialylated gangliosides equally inhibited agglutination of trypsinized rabbit red cells by bovine brain lectin, but not that by the alpha-galactoside-specific antibody from human serum. Results suggested the possibility of endogenous gangliosides acting as cell surface receptors in mediation of brain lectin function.

Alpha-galactoside-binding isolectins from wild jack fruit seed (Artocarpus hirsuta): purification and properties.

Five isolectins with marked specificity for alpha-linked galactose were purified from the wild jack (Artocarpus hirsuta) seeds by affinity chromatography on cross-linked guar gum. They were composed of a glycosylated subunit A (Mr = 16 kDa) and a nonglycosylated subunit B (Mr = 11 kDa) in noncovalent association. The isolectins which eluted as a single peak of Mr 45 kDa on gel filtration in Biogel P-100 and in a TSK G-3000 SW high pressure column, were resolved into five peaks on electrophoresis at pH 4.5. Sodium dodecyl sulphate polyacrylamide gel electrophoreogram of the major isolectin band suggested that the isolectins may be the five possible tetrameric combinations of A and B subunits. The combined isolectins bound only two molecules of 4-methyl umbelliferyl alpha-D-galactoside with a binding constant of 4.75 x 10(4) M-1. The pH optimum of sugar binding was 7.0. The isolectins specifically bound to human IgG and IgA but not to IgM.