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 about 50 to 1500 ms time range, of the binding and dissociation of ribosomal ligands.
Real-time dynamics of ribosome-ligand interaction by time-resolved chemical probing methods
FABBRETTI, Attilio;GIULIODORI, Anna Maria;
2007-01-01
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 about 50 to 1500 ms time range, of the binding and dissociation of ribosomal ligands.File in questo prodotto:
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