Directed Evolution Using Stabilized Bacterial Peptide Display
| Autor: | Tejas Navaratna, Daniel Tresnak, Vivekanandan Subramanian, Lydia Atangcho, Marshall Case, Andrew Min, Mukesh Mahajan, Greg M. Thurber |
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| Rok vydání: | 2019 |
| Předmět: |
medicine.medical_treatment
Sequence (biology) Peptide 010402 general chemistry 01 natural sciences Biochemistry Article Catalysis Colloid and Surface Chemistry Escherichia coli medicine Amino Acids Nuclear Magnetic Resonance Biomolecular chemistry.chemical_classification Protease Chemistry Escherichia coli Proteins General Chemistry Directed evolution Combinatorial chemistry 0104 chemical sciences Amino acid Helix Click chemistry Directed Molecular Evolution Peptides Linker |
| Zdroj: | J Am Chem Soc |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/jacs.9b10716 |
| Popis: | 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 SPEED (Stabilized Peptide Evolution by E. coli Display) 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 (K(d) = 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. |
| Databáze: | OpenAIRE |
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