Selection of a promiscuous minimalist cAMP phosphodiesterase from a library of de novo designed proteins.

ABSTRACT: The ability of unevolved amino acid sequences to become biological catalysts was key to the emergence of life on Earth. However, billions of years of evolution separate complex modern enzymes from their simpler early ancestors. To probe how unevolved sequences can develop new functions, we use ultrahigh-throughput droplet microfluidics to screen for phosphoesterase activity amidst a library of more than one million sequences based on a de novo designed 4-helix bundle. Characterization of hits revealed that acquisition of function involved a large jump in sequence space enriching for truncations that removed >40% of the protein chain. Biophysical characterization of a catalytically active truncated protein revealed that it dimerizes into an α-helical structure, with the gain of function acc

SUBMITTER: Schnettler JD 

PROVIDER: S-EPMC11230910 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature