Co-crystal structure optimization using Isothermal Titration Calorimetry

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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.