Analysis of RNA folding and ligand binding by conventional and high-throughput calorimetry.
Noncoding RNAs serve myriad functions in the cell, but their biophysical properties are not well understood. Calorimetry offers direct and label-free means for characterizing the ligand-binding and thermostability properties of these RNA. We apply two main types of calorimetry--isothermal titration calorimetry (ITC) and differential scanning calorimetry (DSC)--to the characterization of these functional RNA molecules. ITC can describe ligand binding in terms of stoichiometry, affinity, and heat (enthalpy), while DSC can provide RNA stability in terms of heat capacity, melting temperature, and folding enthalpy. Here, we offer detailed experimental protocols for studying such RNA systems with commercially available conventional and high-throughput ITC and DSC instruments.
Department of Biochemistry and Molecular Biophysics, Washington University St Louis School of Medicine, St. Louis, MO, USA. jes513@psu.edu
Joshua E
JE
Sokoloski
Philip C
PC
Bevilacqua
D002151:Calorimetry / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D008024:Ligands; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic
D008024:Ligands; D012313:RNA