Journal
METHODS
Volume 47, Issue 3, Pages 198-205Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2008.09.003
Keywords
ITC; RNA folding; Thermodynamics; Structural rearrangement; Binding
Funding
- NIH [GM-065430, GM-075068]
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Isothermal titration calorimetry (ITC) is a fast and robust method to study the physical basis of molecular interactions. A single well-designed experiment can provide complete thermodynamic characterization of a binding reaction, including K-a, Delta G, Delta H, Delta S and reaction stoichiometry (n). Repeating the experiment at different temperatures allows determination of the heat capacity change (Delta C-P) of the interaction. Modern calorimeters are sensitive enough to probe even weak biological interactions making ITC a very popular method among biochemists. Although ITC has been applied to protein studies for many years, it is becoming widely applicable in RNA biochemistry as well, especially in studies which involve RNA folding and RNA interactions with small molecules, proteins and with other RNAs. This review focuses on best practices for planning, designing and executing effective ITC experiments when one or more of the reactants is an RNA. (c) 2008 Elsevier Inc. All rights reserved.
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