Isolation of fucosyl glycoproteins from human erythrocyte membranes by affinity chromatography using Aleuria aurantia lectin.
Fucosyl glycoproteins were fractionated from a sialoglycoprotein preparation of human erythrocyte membrane by using Aleuria aurantia lectin (AAL) coupled to Sepharose 4B. The affinity eluates were characterized as having high fucose content and significant H activity as measured in terms of N-acetylgalactosamine (GalNAc) incorporation with A1-enzyme and hemagglutination inhibition assay with anti-H sera, and the unadsorbed fractions contained low levels of fucose and were devoid of apparent H activity. Neuraminidase treatment of the material improved the recovery of the affinity eluate. Thus, 66% of the applied asialoglycoprotein was recovered in the eluate, though only 10% of the untreated material was bound and eluted. Moreover, a fucose-rich and H-active fraction was obtained through the affinity chromatography of the previously unbound fraction after neuraminidase treatment. In sodium dodecyl sulfate-gel electrophoresis, the main component of both the unadsorbed and eluted fraction was revealed to be PAS-1 glycoprotein. These results indicate that AAL-Sepharose was effective for isolating fucose-containing compounds from the membrane glycoprotein especially after neuraminidase treatment. The reasons for the appearance of H activity in the affinity eluates are discussed.
D001212:Asialoglycoproteins / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods; D004910:Erythrocyte Membrane / Q000032:analysis*; D005643:Fucose / Q000032:analysis; D006023:Glycoproteins / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D006868:Hydrolysis; D037102:Lectins; D009439:Neuraminidase
D001212:Asialoglycoproteins; D006023:Glycoproteins; D037102:Lectins; D005643:Fucose; D009439:Neuraminidase