Phage-assisted evolution of allosteric protein switches - PubMed
3 hours ago
- #allosteric protein switches
- #optogenetic switches
- #phage-assisted evolution
- Researchers introduced a phage-assisted evolution platform for optimizing allosteric protein switches in vivo, applying opposing selection pressures to enhance activity and switchability.
- The platform uses retron-based recombineering to introduce diverse mutations (point mutations, insertions, deletions) and explore fitness landscapes, with application to transcription factor AraC yielding near-binary optogenetic switches with ~1000-fold dynamic range.
- Long-read sequencing across selection cycles tracked evolving variant pools, revealing adaptive trajectories and context-dependent residue interactions, identifying linker mutations that promote α-helix extension at the sensor-effector junction to enhance conformational coupling.