Chapter title |
Isothermal Titration Calorimetry: Assisted Crystallization of RNA–Ligand Complexes
|
---|---|
Chapter number | 9 |
Book title |
Nucleic Acid Crystallography
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-2763-0_9 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2762-3, 978-1-4939-2763-0
|
Authors |
Cyrielle Da Veiga, Joelle Mezher, Philippe Dumas, Eric Ennifar, Veiga, Cyrielle Da, Mezher, Joelle, Dumas, Philippe, Ennifar, Eric |
Abstract |
The success rate of nucleic acids/ligands co-crystallization can be significantly improved by performing preliminary biophysical analyses. Among suitable biophysical approaches, isothermal titration calorimetry (ITC) is certainly a method of choice. ITC can be used in a wide range of experimental conditions to monitor in real time the formation of the RNA- or DNA-ligand complex, with the advantage of providing in addition the complete binding profile of the interaction. Following the ITC experiment, the complex is ready to be concentrated for crystallization trials. This chapter describes a detailed experimental protocol for using ITC as a tool for monitoring RNA/small molecule binding, followed by co-crystallization. |
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