title: Binding specificities of lectins to immobilized glycoproteins and oligosaccharides differ from those of immobilized lectins to oligosaccharides.
abstract: The carbohydrate-binding specificities of lectins in solution to glycoproteins and neoglycolipids immobilized on a solid phase were analyzed in order to establish a simple, rapid method for structural analysis of the carbohydrate moieties of small amounts of individual glycoproteins blotted on membrane. Eight glycoproteins containing typical O-linked tetrasaccharides or a series of typical N-linked oligosaccharides of the high-man-nose type, hybrid type, and complex type and 6 neoglycoproteins containing mono- or di-saccharides were dot blotted on membranes and the membranes were then reacted with 8 kinds of horseradish peroxidase-conjugated lectins before and after heat treatment. Neoglycolipids containing the glycoprotein-derived oligosaccharides immobilized on a thin layer chromatography plate were also reacted with lectins. The heat treatment of the membrane increased lectin reactivity toward the glycoproteins. The carbohydrate-binding behavior of lectins, Phaseolus vulgaris erythroagglutinin, wheat germ agglutinin, and concanavalin A in solution toward glycoproteins and neoglycolipids immobilized on a solid phase differed from that of immobilized lectins toward oligosaccharides in solution. This difference should be noted in lectin detection of specific carbohydrates of individual glycoproteins on membrane.
authors.affiliation: Department of Applied Chemistry, Tokai University, Kanagawa, Japan.
authors.forename: W
authors.initials: W
authors.lastname: Tang
authors.forename: T
authors.initials: T
authors.lastname: Miura
authors.forename: M
authors.initials: M
authors.lastname: Nakata
authors.forename: N
authors.initials: N
authors.lastname: Kojima
authors.forename: T
authors.initials: T
authors.lastname: Mizuochi
mesh_terms: D006023:Glycoproteins / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D006023:Glycoproteins; D037102:Lectins; D009844:Oligosaccharides