title: Activation of RNase L by 2',5'-oligoadenylates. Kinetic characterization.
abstract: Ribonuclease L (RNase L), the 2',5'-oligoadenylate-dependent ribonuclease, is one of the cellular antiviral systems with enhanced activity in the presence of interferon. A reaction scheme has been developed to model the sequence of steps necessary for the activation of RNase L (Cole, J. L., Carroll, S. S., Blue, E. S., Viscount, T., and Kuo, L. C. (1997) J. Biol. Chem. 272, 19187-19192). The model comprises three sequential binding steps: the binding of activator to enzyme monomer, the subsequent dimerization of the activated monomer to form the active enzyme dimer, followed by the binding of substrate prior to catalysis. The model is used to evaluate the activation of RNase L by several synthetic analogs of the native activator. The 5'-phosphate of the activator has been determined to be an important structural determinant for the efficient activation of RNase L, and its loss caused a loss of activator affinity of 2-3 orders of magnitude. The length of activator is not an important determinant of activator potency for the activator analogs examined. The specific activity of the enzyme under conditions of saturation of activator binding and complete dimerization of the activated monomers varies only by about a factor of 3 for the activators examined, indicating that once dimerized in the presence of any of these activators, the enzyme exhibits a similar catalytic activity.
authors.affiliation: Department of Antiviral Research, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
authors.forename: S S
authors.initials: SS
authors.lastname: Carroll
authors.forename: J L
authors.initials: JL
authors.lastname: Cole
authors.forename: T
authors.initials: T
authors.lastname: Viscount
authors.forename: J
authors.initials: J
authors.lastname: Geib
authors.forename: J
authors.initials: J
authors.lastname: Gehman
authors.forename: L C
authors.initials: LC
authors.lastname: Kuo
mesh_terms: D000227:Adenine Nucleotides / Q000494:pharmacology*; D019281:Dimerization; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D009843:Oligoribonucleotides / Q000494:pharmacology*; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000227:Adenine Nucleotides; D009843:Oligoribonucleotides; C023505:2',5'-oligoadenylate; D012260:Ribonucleases