A solid-phase radio-binding assay for the characterization of lectin recognition.
A rapid and quantitative method for characterizing lectin specificity is presented. This assay is analogous to solid-phase radioimmune assays and utilizes the property of irreversible binding of proteins to vinyl microtiter plates. The lectins phytohemagglutinin and concanavalin A bind to vinyl and retain their biological properties. Iodinated fetuin and immunoglobulin G are both bound by the immobilized lectins. Competing amounts of ligands presented at the same time as the iodinated glycoproteins are shown to reduce binding to the immobilized lectins. Conversely, glycoprotein ligands immobilized to vinyl retain their ability to be recognized and bind lectins. The solid-phase binding assay can be used to characterize the ligand-binding specificity of lectins. For example, the pattern of glycoprotein inhibition of the binding of iodinated gorgonian lectin from Leptogorgia virgulata to insolubilized thyroglobulin is virtually identical to the pattern reported previously using hemagglutination inhibition. The solid-phase radio-binding assay is rapid, reproducible, and sensitive to nanogram quantities of ligand added. Most importantly, it provides quantitative information on lectin recognition.
D000818:Animals; D001212:Asialoglycoproteins; D001667:Binding, Competitive; D002417:Cattle; D006020:Glycopeptides / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D037102:Lectins / Q000032:analysis*; D011485:Protein Binding; D011869:Radioligand Assay; D012995:Solubility; D013954:Thyroglobulin / Q000031:analogs & derivatives / Q000378:metabolism
D001212:Asialoglycoproteins; D006020:Glycopeptides; D037102:Lectins; C019149:asialothyroglobulin; D013954:Thyroglobulin