title: Purification of mammalian ribonuclease using immobilized human ribonuclease inhibitor.
abstract: Affinity chromatography on immobilized ribonuclease (RNase) inhibitor was developed for purification of mammalian RNase. Human placental RNase inhibitor was conjugated to CNBr-activated Sepharose in the presence of dithiothreitol. About 80% of the immobilized RNase inhibitor was capable of binding bovine pancreatic RNase A. The bound RNase A was eluted with 3 M NaCl at pH 5.0. Two 25-kDa and 18-kDa RNases, which were obtained from human liver using a cellulose phosphate column, were bound to the immobilized RNase inhibitor and recovered in a pure and active form after treatment of the resin with p-hydroxymercuribenzoate. These enzymes were considered to be nonsecretory-type RNases with different sugar contents.
authors.affiliation: Department of Legal Medicine, Fukui Medical School, Japan.
authors.forename: D
authors.initials: D
authors.lastname: Nadano
authors.forename: T
authors.initials: T
authors.lastname: Yasuda
authors.forename: H
authors.initials: H
authors.lastname: Takeshita
authors.forename: K
authors.initials: K
authors.lastname: Sawazaki
authors.forename: K
authors.initials: K
authors.lastname: Kishi
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D002411:Cation Exchange Resins / Q000378:metabolism; D002417:Cattle; D002482:Cellulose / Q000031:analogs & derivatives / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010920:Placenta / Q000201:enzymology; D010926:Placental Hormones / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D012260:Ribonucleases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D002411:Cation Exchange Resins; D007527:Isoenzymes; D010926:Placental Hormones; C022052:placental ribonuclease inhibitor; D012313:RNA; D002482:Cellulose; C008881:phosphocellulose; D012260:Ribonucleases; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase