title: First demonstration of lactoribonuclease, a ribonuclease from bovine milk with similarity to bovine pancreatic ribonuclease.
abstract: The isolation of a ribonuclease designated lactoribonuclease, with a molecular weight and an N-terminal amino acid sequence identical to those of bovine pancreatic ribonuclease, was first reported from bovine milk. After removal of globulin from acid whey by precipitation with 1.8 M (NH4)2SO4, (NH4)2SO4 was added to attain a concentration of 3.6 M. Adsorption on the ion exchanger CM-Sepharose and subsequently on Mono S by fast protein liquid chromatography yielded pure lactoribonuclease. The enzyme, like pancreatic ribonuclease, was most active at pH 7.5 with yeast transfer RNA (tRNA) as substrate. Lactoribonuclease and pancreatic ribonuclease showed a strong preference for poly(C) over poly(U). However, pancreatic ribonuclease did so with a higher specific activity, suggesting that the two ribonucleases are not identical. No inhibitory effect was shown by either lactoribonuclease or pancreatic ribonuclease toward poly (A) and poly (G). The effect of lactoribonuclease and pancreatic ribonuclease on tRNA increased with the concentration of tRNA. Lactoribonuclease inhibited cell-free translation in a rabbit reticulocyte lysate system with an IC50 of 3.5 nM while the corresponding IC50 for pancreatic ribonuclease was 0.09 nM.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, NT.
authors.forename: X Y
authors.initials: XY
authors.lastname: Ye
authors.forename: T B
authors.initials: TB
authors.lastname: Ng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D004722:Endoribonucleases / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D008892:Milk / Q000201:enzymology*; D008969:Molecular Sequence Data; D012343:RNA, Transfer / Q000378:metabolism; D012259:Ribonuclease, Pancreatic / Q000032:analysis* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D012343:RNA, Transfer; D004722:Endoribonucleases; C415668:lactoribonuclease; D012259:Ribonuclease, Pancreatic