記録された日時: March 24 2015

Duration: 39 minutes 00 seconds

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.

Table of contents
1. Untitled
02:06
2. Microcalorimetry as a Tool for Structural Biology
00:28
3. Brief Introduction to ITC
01:11
4. Benefits from ITC
01:54
5. Common Issues in Crystallization of Complexes
01:38
6. Example #1
00:34
7. HIV-1 Genomic RNA Dimerization Initiation Site
00:54
8. HIV-1 Genomic RNA Dimerization Initiation Site
01:02
9. Distinction Between Specific and Unspecific Binding
02:04
10. Natrix HT Screen
00:41
11. Example #2
00:21
12. HIV-1 Reverse Transcriptase
01:25
13. Graph
01:28
14. Graph
01:10
15. Graph
00:50
16. Primer/Template
02:20
17. Example #3
00:16
18. B Sliding Clamp
01:26
19. Wolff et al, J Med Chem 2014
00:21
20. Example #4
00:20
21. Riboswitches
00:51
22. Goal of Study
00:42
23. Graph
01:35
24. Conclusions
00:08
25. ITC-assisted Crystalization
01:02
26. Biophysics and Structural Biology
01:13
27. Thank you for your attention
08:54
28. Contact Information
02:06
29. Find out more about the MicroCal ITC range
00:00