title: Heterogeneity in the glycosylation pattern of human pancreatic ribonuclease.
abstract: Different molecular forms of ribonuclease were isolated from fresh human pancreas obtained from healthy transplant donors. The purification procedure consists of the preparation of an acetone powder, followed by (NH4)2SO4 precipitation and two chromatography steps (cationic exchange and reversed-phase). Protein bands in gel electrophoresis with RNAase activity were monitored using a negative-staining zymogram technique. Several glycosylated enzyme forms with apparent molecular masses ranging from 14 to 40 kDa were separated. Peptides containing the three Asn-Xaa-Thr/Ser acceptor sites for glycosylation were isolated and analysed. The site with Asn-34 was almost completely glycosylated, while the sites with Asn-76 and Asn-88 had carbohydrate in about half and a minor part of the molecules, respectively. The carbohydrate compositions of the glycopeptides are different from those of the same gene product isolated from human urine. C-Terminal threonine was present in part of the molecules, indicating partial degradation by carboxypeptidase.
authors.affiliation: Institut de Biologia Fonamental V. Villar Palasí, Facultat de Ciències, Universitat Autònoma de Barcelona, Bellaterra, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Ribó
authors.forename: J J
authors.initials: JJ
authors.lastname: Beintema
authors.forename: M
authors.initials: M
authors.lastname: Osset
authors.forename: E
authors.initials: E
authors.lastname: Fernández
authors.forename: J
authors.initials: J
authors.lastname: Bravo
authors.forename: R
authors.initials: R
authors.lastname: De Llorens
authors.forename: C M
authors.initials: CM
authors.lastname: Cuchillo
mesh_terms: D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D012260:Ribonucleases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013912:Threonine / Q000032:analysis
chemical_list: D002241:Carbohydrates; D013912:Threonine; D012260:Ribonucleases