Metal-responsive regulation of enzyme catalysis using genetically encoded chemical switches
Abstract
Dynamic control over protein function is a central challenge in synthetic biology. To address this challenge, we describe the development of an integrated computational and experimental workflow to incorporate a metal-responsive chemical switch into proteins. Pairs of bipyridinylalanine (BpyAla) residues are genetically encoded into two structurally distinct enzymes, a serine protease and firefly luciferase, so that metal coordination biases the conformations of these enzymes, leading to reversible control of activity. Computational analysis and molecular dynamics simulations are used to rationally guide BpyAla placement, significantly reducing experimental workload, and cell-free protein synthesis coupled with high-throughput experimentation enable rapid prototyping of variants. Ultimately, this strategy yields enzymes with a robust 20-fold dynamic range in response to divalent metal salts over 24 on/off switches, demonstrating the potential of this approach. We envision that this strategy of genetically encoding chemical switches into enzymes will complement other protein engineering and synthetic biology efforts, enabling new opportunities for applications where precise regulation of protein function is critical.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Apr 06, 2022
- Source ID
- 10.1038/s41467-022-29239-y
Entities
People
- Andrew C Hunt
- Benoît Roux
- Bingqing Liu
- Jared C Lewis
- Kosuke Seki
- Michael C Jewett
- Yasmine S. Zubi
- Ying Li
Organizations
- Army Research Office
- David and Lucile Packard Foundation
- National Institute of General Medical Sciences
- The Camille and Henry Dreyfus Foundation
- United States Department of Health and Human Services