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1. Introduction
2. Goal of described researches
3. How do small molecules bind proteins?
4. Flexible regions in small molecule binding (1)
5. P-loop structures
6. Flexible regions in small molecule binding (2)
7. Methods- phage display
8. Phage display technology
9. Biopanning
10. Phage display - advantages
11. Evaluating library diversity
12. What do we mean by diversity?
13. An intuitive analytical expression of diversity
14. Example (1)
15. Example (2)
16. Example (3)
17. Example (4)
18. The probability to select the same peptide twice
19. RELIC
20. Sequence diversity of proteomes and libraries
21. 12mer libraries with same amino acid distribution
22. Identifying motifs
23. Using diverse populations to identify ATP-BP
24. Consensus sequences as clues to protein function
25. Peptide libraries displayed on phage surface
26. Two ATP substrates used
27. For this to work you need lots of peptides
28. How many P-loops did we select?
29. P-loops are censored from the unselected library!
30. Ratio of ATP-selected peptides
31. Phosphoenolpyruvate carboxykinase
32. Can the method identify ATP-BP?
33. Taxol
34. Taxol- targets and activities
35. Immobilization of taxol through biotinylation
36. One segment of tubulin with high similarity to taxol
37. An alternative means of displaying the results
38. Bcl-3
39. Selected peptides and the flexible loop of Bcl-3
40. Taxol binds to the flexible loop of Bcl-3
41. ELISA data
42. CD spectra
43. Taxol - Bcl-2 interaction
44. Taxol-selected peptides and mdr1/mdr4
45. Taxol binds to loops on the surface of mdr2
46. Taxol's targets
47. Summary
48. Thank you

Topics Covered

• How do you identify the molecular target of a small molecule drug?
• Flexible regions are key players
• P-loop of ATP-binding proteins is an important example
• Can display libraries of flexible peptides on proteins select those that bind to the ligand
• Selection is through an affinity process called biopanning
• Success depends on the diversity of the library
• Diversity depends on how many different peptides are in the library and their relative abundance
• RELIC can be used to calculate the diversity of a library from the sequences of ~100 peptides
• Identification of motifs from a population is easy if the motif is very strong
• Taxol-binding peptides were also identified by phage display and those peptides were used to identify the position on tubulin where taxol is bound and to identify Bcl-2 as a taxolbinding protein
• Significant data from a variety of sources argue that the taxol-Bcl-2 interaction is important to the clinical efficacy of taxol

Links

Series:

• Chemical Biology

Categories:

• Biochemistry
• Methods

Talk Citation

Makowski, L. (2008, August 14). Mapping small molecule binding sites with phage display technology [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved April 16, 2024, from https://hstalks.com/bs/948/.
Export Citation (RIS)

Publication History

• Published on August 14, 2008
• Archived on August 13, 2020

Financial Disclosures

• Dr. Lee Makowski has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.