Cryoenzymology of the hammerhead ribozyme

Andrew L. Feig, Glenyss E. Ammons, Olke C. Uhlenbeck*

*Corresponding author for this work

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

The technique of cryoenzymology has been applied to the hammerhead ribozyme in an attempt to uncover a structural rearrangement step prior to cleavage. Several cryosolvents were tested and 40% (v/v) methanol in water was found to perturb the system only minimally. This solvent allowed the measurement of ribozyme activity between 30 and -33 °C. Eyring plots are linear down to -27 °C, but a drastic reduction in activity occurs below this temperature. However, even at extremely low temperatures, the rate is still quite pH dependent, suggesting that the chemical step rather than a structural rearrangement is still rate-limiting. The nonlinearity of the Eyring plot may be the result of a transition to a cold-denatured state or a glassed state.

Original languageEnglish (US)
Pages (from-to)1251-1258
Number of pages8
JournalRNA
Volume4
Issue number10
DOIs
StatePublished - Oct 1 1998

Keywords

  • Conformational change
  • Kinetics
  • Temperature dependence

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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