Schistosoma mansoni: radiometric assay of lectin binding specificities of the major egg glycoprotein and its carbohydrate-rich fragment.
The binding by lectins of the Schistosoma mansoni major egg glycoprotein and of a carbohydrate-rich fragment which is serologically cross-reactive with it was studied. The major egg glycoprotein was purified from a crude soluble egg antigen by a succession of affinity chromatography procedures on concanavalin A-sepharose and by ion-exchange chromatography. The carbohydrate-rich fragment was isolated by ultrafiltration of the crude glycoprotein fraction initially obtained from the crude soluble egg antigens. The major egg glycoprotein and the carbohydrate-rich fragment contain 77 and 92.5% carbohydrate, respectively. When radioiodinated and run on SDS-polyacrylamide gel electrophoresis, each of them exhibited a single peak with respective Rf values of 0.33 and 1.0, and their respective molecular weights were 70K and 10-13K. The binding of the radioiodinated major egg glycoprotein and the carbohydrate-rich fragment by peanut agglutinin, Ricinus communis agglutinin-60, wheat germ agglutinin, and lotus agglutinin was studied by double diffusion in agar, and by a radiometric solid-phase assay in which the lectins were used to coat microtiter plates. The latter assay was employed to determine the specificity of the binding by inhibition with the specific sugars. Both the major egg glycoprotein and the carbohydrate-rich fragment bound specifically to concanavalin A columns as indicated by their isolation procedure. They also bound specifically to peanut agglutinin, R. communis agglutinin 60, and lotus agglutinin, while binding by wheat germ agglutinin appeared not to be specific.(ABSTRACT TRUNCATED AT 250 WORDS)
D000116:Acetylgalactosamine / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis*; D004527:Egg Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D005643:Fucose / Q000032:analysis; D005690:Galactose / Q000032:analysis; D005947:Glucose / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D037102:Lectins; D008358:Mannose / Q000032:analysis; D019887:Peanut Agglutinin; D010446:Peptide Fragments / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D037121:Plant Lectins; D012550:Schistosoma mansoni / Q000032:analysis*; D014909:Wheat Germ Agglutinins
D002241:Carbohydrates; D004527:Egg Proteins; D006023:Glycoproteins; D037102:Lectins; D019887:Peanut Agglutinin; D010446:Peptide Fragments; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014909:Wheat Germ Agglutinins; D005643:Fucose; D005947:Glucose; D000116:Acetylgalactosamine; D008358:Mannose; D005690:Galactose