Studies of lectin-carbohydrate interactions by quantitative affinity chromatography: systems with galactose and ovalbumin as saccharidic ligand.
The potential of affinity chromatography for characterizing lectin-carbohydrate interactions is investigated. First, the effect of galactose on the chromatographic behavior of Ricinus communis phytohemagglutinin on Sepharose 4B is used to establish that quantitative affinity chromatography on polysaccharide matrices affords an unequivocal means of characterizing the interactions of lectins with monosaccharides in solution. Second, a method of characterizing lectin-glycoprotein interactions by affinity chromatography is illustrated in an experimental study with Sephadex G-50 as affinity matrix for examination of the interaction between concanavalin A and ovalbumin. Third, although no general solution to the problem of ligand multivalency in quantitative affinity chromatography has been found, an experimental protocol has been devised for the situation in which the partitioning solute (lectin) is univalent.
Hogg
P J
PJ
Department of Biochemistry, University of Queensland, St. Lucia, Australia.
Winzor
D J
DJ
D001665:Binding Sites; D002241:Carbohydrates / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D005690:Galactose / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification; D037102:Lectins / Q000302:isolation & purification*; D008024:Ligands; D010047:Ovalbumin / Q000302:isolation & purification
D002241:Carbohydrates; D006023:Glycoproteins; D037102:Lectins; D008024:Ligands; D010047:Ovalbumin; D005690:Galactose