title: Characterization of human arylsulfatase A glycans.
abstract: Despite numerous studies on arylsulfatase A, the structure of the glycans present in each of its two subunits has not been determined. This is important because the carbohydrate component of human arylsulfatase A synthesized in tumor tissues and transformed cells has been shown to undergo apparent changes. This study elucidates some of their major features. Glycan chain analysis of native and deglycosylated arylsulfatase A as well as its subunits was performed with the use of a Glycan Differentiation Kit and lectin affinity chromatography. Each of the two subunits of arylsulfatase A from placenta, separated electrophoretically on polyacrylamide gel in reducing conditions, reacted with digoxigenin-labelled Galantus nivalis agglutinin and Aleuria aurantia agglutinin, while those from liver enzyme reacted with the former only. The subunits of both enzymes did not react with Sambucus nigra, Maakia amuriensis, Datura stramonium or Peanut agglutinin. Deglycosylation of arylsulfatase A with peptide N-glycosidase F and endo-beta-N-acetylglucosaminidase F resulted in complete cleavage of its carbohydrate component from each subunit. Their molecular weights decreased by 3 kDa. Neuraminidase treatment of the enzyme from liver and placenta followed by isoelectrofocusing separation showed the presence of sialylated forms which constituted a small percentage of total enzyme activity. Placental arylsulfatase A became bound to Lens culinaris agglutinin agarose, while no interaction with Ricinus communis or Griffonia simplicifolia agglutinin agarose was observed. The study shows that both subunits of arylsulfatase A from human placenta possess two high mannose/hybrid type glycans as major structures, with at least one 6-O-L-fucose bound to the innermost N-acetylglucosamine on each.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Institute of Medical Biochemistry, Jagiellonian University, Krakow, Poland.
authors.forename: P
authors.initials: P
authors.lastname: Laidler
authors.forename: A
authors.initials: A
authors.lastname: Litynska
authors.forename: M
authors.initials: M
authors.lastname: Galka-Walczak
authors.forename: B
authors.initials: B
authors.lastname: Wojczyk
mesh_terms: D002240:Carbohydrate Sequence; D002553:Cerebroside-Sulfatase / Q000737:chemistry*; D006801:Humans; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology; D011134:Polysaccharides / Q000737:chemistry*
chemical_list: D011134:Polysaccharides; D002553:Cerebroside-Sulfatase