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Directed Evolution Using Stabilized Bacterial Peptide Display
- Source :
- Journal of the American Chemical Society; January 2020, Vol. 142 Issue: 4 p1882-1894, 13p
- Publication Year :
- 2020
-
Abstract
- Chemically stabilized peptides have attracted intense interest by academics and pharmaceutical companies due to their potential to hit currently “undruggable” targets. However, engineering an optimal sequence, stabilizing linker location, and physicochemical properties is a slow and arduous process. By pairing non-natural amino acid incorporation and cell surface click chemistry in bacteria with high-throughput sorting, we developed a method to quantitatively select high affinity ligands and applied the Stabilized Peptide Evolution by E. coliDisplay technique to develop disrupters of the therapeutically relevant MDM2-p53 interface. Through in situ stabilization on the bacterial surface, we demonstrate rapid isolation of stabilized peptides with improved affinity and novel structures. Several peptides evolved a second loop including one sequence (Kd= 1.8 nM) containing an i, i+4 disulfide bond. NMR structural determination indicated a bent helix in solution and bound to MDM2. The bicyclic peptide had improved protease stability, and we demonstrated that protease resistance could be measured both on the bacterial surface and in solution, enabling the method to test and/or screen for additional drug-like properties critical for biologically active compounds.
Details
- Language :
- English
- ISSN :
- 00027863 and 15205126
- Volume :
- 142
- Issue :
- 4
- Database :
- Supplemental Index
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Periodical
- Accession number :
- ejs51901502
- Full Text :
- https://doi.org/10.1021/jacs.9b10716