title: Affinity chromatography of branched oligosaccharides in rat liver beta-glucuronidase.
abstract: Rat liver microsomal and lysosomal beta-glucuronidase-derived glycopeptides were obtained by extensive Pronase digestion followed by N-[14C]acetylation and desialylation by neuraminidase treatment. These glycopeptides were studied by sequential chromatography on lectin-affinity columns such as concanavalin A, lentil lectin, Phaseolus vulgaris erythroagglutinin, Ricinus communis agglutinin I, Triticum vulgaris agglutinin, Glycine max agglutinin and Ulex europaeus agglutinin. Using serial lectin affinity chromatography approach combined with neuraminidase treatment allowed us to show the unexpected presence of complex tri- and/or tetraantennary type glycans (40.8 and 17.0% for microsomal and lysosomal enzyme, respectively). Moreover, the application of neuraminidase treatment revealed that complex biantennary type glycans, present on lysosomal beta-glucuronidase, are almost fully sialylated while the same type of glycans present on microsomal enzyme do not contain sialic acid. Furthermore, the results obtained confirmed that microsomal and lysosomal beta-glucuronidases possess high mannose and/or hybrid type glycans (19.6 and 36.6%, respectively), and complex biantennary type glycans (38.9 and 46.4%, respectively).
authors.affiliation: Department of Animal Physiology, Institute of Zoology, Jagiellonian University, Kraków, Poland. hoja@zuk.iz.uj.edu.pl
authors.forename: D
authors.initials: D
authors.lastname: Hoja-Lukowicz
authors.forename: A
authors.initials: A
authors.lastname: Lityńska
authors.forename: B S
authors.initials: BS
authors.lastname: Wójczyk
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D005966:Glucuronidase / Q000737:chemistry* / Q000302:isolation & purification; D006020:Glycopeptides / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry; D008099:Liver / Q000378:metabolism*; D008247:Lysosomes / Q000737:chemistry; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*; D011402:Pronase; D051381:Rats
chemical_list: D006020:Glycopeptides; D037102:Lectins; D009844:Oligosaccharides; D005966:Glucuronidase; D011402:Pronase