Isothermal Titration Calorimetry Analysis of a Cooperative Riboswitch Using an Interdependent-Sites Binding Model

Methods Mol Biol. 2023:2568:53-73. doi: 10.1007/978-1-0716-2687-0_5.


Isothermal titration calorimetry (ITC) is a powerful biophysical tool to characterize energetic profiles of biomacromolecular interactions without any alteration of the underlying chemical structures. In this protocol, we describe procedures for performing, analyzing, and interpreting ITC data obtained from a cooperative riboswitch-ligand interaction.

Keywords: Cooperativity; Ligand binding; Nonequivalent-interdependent-sites model; Parabolic thermogram; Riboswitch; Stoichiometry; Thermodynamics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calorimetry / methods
  • Ligands
  • Protein Binding
  • Riboswitch*
  • Thermodynamics


  • Ligands
  • Riboswitch