title: Purification and mode of action of a microsomal exoribonuclease from rat liver.
abstract: An exoribonuclease has been purified nearly to homogeneity from rat liver microsomes and its mode of action and general properties were studied. The molecular weight values for the enzyme, as estimated by gel filtration and SDS-polyacrylamide gel electrophoresis, were 88 000 and 92 000, respectively. The enzyme produced, via a processive mechanism Ado5'P as the only product from poly(A). The results of the hydrolysis of 4 S (Ado5'P)n and (Ado3'P)n by the exoribonuclease with or without alkaline phosphatase and the inhibition of the enzymatic activity by oligonucleotides having a 3'-phosphate group in the 3'-terminus suggested that the degradation proceeds in the 3' to 5' direction. These findings were confirmed by the analysis of hydrolyzed products of various oligoadenylates and Ado3'PUrdPGuo and by the comparison of the rates of hydrolysis of (Ado3'P)2Ado by the enzyme in the presence of varying amounts of (Ado3'P)3. Mg2+ was required for the enzymatic activity, and Mn2+ partially substituted for Mg2+. The activity of the enzyme was stimulated by K+ and spermine.
authors.forename: H
authors.initials: H
authors.lastname: Kumagai
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
authors.forename: N
authors.initials: N
authors.lastname: Ozaki
authors.forename: K
authors.initials: K
authors.lastname: Igarashi
authors.forename: S
authors.initials: S
authors.lastname: Hirose
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005092:Exonucleases / Q000302:isolation & purification*; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D011061:Poly A / Q000378:metabolism; D011066:Poly C / Q000378:metabolism; D011072:Poly U / Q000378:metabolism; D051381:Rats; D012259:Ribonuclease, Pancreatic / Q000378:metabolism
chemical_list: D011061:Poly A; D011072:Poly U; D011066:Poly C; D008345:Manganese; D005092:Exonucleases; D012259:Ribonuclease, Pancreatic; D000469:Alkaline Phosphatase; D008274:Magnesium