Determination of kinetic parameters for hammerhead and hairpin ribozymes.
The application of conventional enzymological methods to the study of hairpin and hammerhead ribozymes has led to valuable insights into the mechanisms by which these small RNAs catalyze phosphodiester cleavage and ligation reactions. Here, protocols are presented for measuring rate constants for simple cleavage and ligation reactions mediated by minimal hammerhead and hairpin ribozymes under standard experimental conditions. Information is also provided to help researchers recognize and interpret more complex reaction kinetics that can be observed for ribozyme-sequence variants under a variety of reaction conditions.
Fedor
Martha J
MJ
Department of Molecular Biology and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, San Diego, CA, USA.
D002384:Catalysis; D007202:Indicators and Reagents; D007700:Kinetics; D016337:RNA, Catalytic / Q000378:metabolism*; D013379:Substrate Specificity
D007202:Indicators and Reagents; D016337:RNA, Catalytic; C418481:hairpin ribozyme; C427204:hammerhead ribozyme