| 00:00:00 | Untitled |
| 00:02:06 | Microcalorimetry as a Tool for Structural Biology |
| 00:02:34 | Brief Introduction to ITC |
| 00:03:45 | Benefits from ITC |
| 00:05:39 | Common Issues in Crystallization of Complexes |
| 00:07:17 | Example #1 |
| 00:07:51 | HIV-1 Genomic RNA Dimerization Initiation Site |
| 00:08:45 | HIV-1 Genomic RNA Dimerization Initiation Site |
| 00:09:47 | Distinction Between Specific and Unspecific Binding |
| 00:11:51 | Natrix HT Screen |
| 00:12:32 | Example #2 |
| 00:12:53 | HIV-1 Reverse Transcriptase |
| 00:14:18 | Graph |
| 00:15:46 | Graph |
| 00:16:56 | Graph |
| 00:17:46 | Primer/Template |
| 00:20:06 | Example #3 |
| 00:20:22 | B Sliding Clamp |
| 00:21:48 | Wolff et al, J Med Chem 2014 |
| 00:22:09 | Example #4 |
| 00:22:29 | Riboswitches |
| 00:23:20 | Goal of Study |
| 00:24:02 | Graph |
| 00:25:37 | Conclusions |
| 00:25:45 | ITC-assisted Crystalization |
| 00:26:47 | Biophysics and Structural Biology |
| 00:28:00 | Thank you for your attention |
| 00:36:54 | Contact Information |
| 00:39:00 | Find out more about the MicroCal ITC range |
The success rate for protein- or RNA/DNA-ligand 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/DNA- or protein-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.
Here we describe how ITC can be used as a tool for monitoring complex formation, followed by co-crystallization.