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1. Introduction
2. Libraries of de novo amino acid sequences
3. Designed libraries of folded proteins
4. Amphipathic alpha helix and beta strand
5. Binary patterned 4-helix bundle
6. Suggested sequence for alpha-helix
7. Genetic code for the suggested sequence
8. Rational design and combinatorial approach
9. Cloning the library into E.coli
10. Example of several sequences from the library
11. Combinatorial libraries of sequences
12. Well folded "native-like" structures
13. Most of the proteins were molten globules
14. Design: 74 amino acid, 14 residues/helix
15. The first and the second generation libraries
16. Stability of the 1st and 2nd generation libraries
17. NOESY spectra
18. NMR for proteins from the unselected library
19. Experimentally determent structure
20. Superposition of low energy structures
21. Polar/nonpolar periodicity of de novo 4-helix bundle
22. Sequence of S-824
23. Functional proteins in unselected binary libraries
24. Heme binding
25. Functions of heme proteins
26. Peroxidase reaction
27. Screen for peroxidase activity
28. Peroxidase activity by oxidation of ABTs
29. Catalysis without cofactors?
30. Hydrolysis of p-nitrophenyl acetate by S-824
31. Esterase activity kcat/kuncat = 8700
32. A binary code for protein design
33. Binary patterning of alpha-helix and beta strand
34. A designed beta sheet template
35. Binary patterned beta-sheet sequences
36. Alpha-helix versus beta-sheet
37. De novo beta-sheet proteins
38. Binary patterns in natural sequences
39. Engineering monomeric beta-sheet proteins
40. Chromatography and circular dichroism
41. Summary for binary code for protein design
42. Biomaterials
43. Binary patterned design for beta-sheet proteins
44. Model of graphite surface and AFM image
45. Acknowledges

Topics Covered

• Novel proteins can be devised using either rational/computational approaches or random/combinatorial methods
• Description of a hybrid approach that combines rational and combinatorial methods: libraries of novel proteins
• Combinatorial diversity
• Random vs. focused libraries
• Amphiphilic alpha-helices and beta-strands
• Binary patterning of polar and non-polar amino acids
• Genetic encoding of combinatorial mixtures of polar and non-polar amino acids
• Design of libraries of novel 4-helix bundles
• Library construction and expression in vivo
• Dynamic vs. wellordered protein structures
• Second generation libraries
• Biophysical characterization
• Solution structure of native-like 4-helix bundle proteins from designed libraries
• Functionally active proteins including cofactor binding proteins, peroxidases and esterases
• Designed libraries of beta sheet proteins: amyloid-like fibrils, monomeric beta-sheet proteins and novel biomaterials

Links

Series:

• Protein Folding, Aggregation and Design

Categories:

• Biochemistry
• Methods

Talk Citation

Hecht, M. (2007, October 1). Novel proteins from designed combinatorial libraries [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved November 23, 2024, from https://doi.org/10.69645/UTMA5950.
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Publication History

• Published on October 1, 2007
• Archived on April 3, 2017

Financial Disclosures

• Prof. Michael Hecht has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.