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Real-Time Dynamics of Ribosome-Ligand Interaction by Time-Resolved Chemical Probing Methods

  • Attilio Fabbretti
  • , Pohl Milon
  • , Anna Maria Giuliodori
  • , Claudio O. Gualerzi
  • , Cynthia L. Pon
  • University of Camerino

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

13 Scopus citations

Abstract

Three protocols to perform time-resolved in situ probing of rRNA are described. The three methods (chemical modification with DMS and rRNA backbone cleavage by hydroxyl radicals generated by either K-peroxonitrite or Fe(II)-EDTA) make use of a quench-flow apparatus and exploit reactions that are faster than the interactions of ribosomal subunits with their ligands. These methods allow the investigation of the path and dynamics, in a ≅ 50 to 1500 ms time range, of the binding and dissociation of ribosomal ligands.

Original languageEnglish
Title of host publicationTranslation Initiation
Subtitle of host publicationReconstituted Systems and Biophysical Methods
PublisherAcademic Press Inc.
Pages45-58
Number of pages14
ISBN (Print)9780123739698
DOIs
StatePublished - 2007
Externally publishedYes

Publication series

NameMethods in Enzymology
Volume430
ISSN (Print)0076-6879

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