Data from: De novo active sites for resurrected Precambrian enzymes

Protein engineering studies often suggest the emergence of completely new
enzyme functionalities to be highly improbable. However, enzymes likely
catalysed many different reactions already in the last universal common
ancestor. Mechanisms for the emergence of completely new active sites must
therefore either plausibly exist or at least have existed at the
primordial protein stage. Here, we use resurrected Precambrian proteins as
scaffolds for protein engineering and demonstrate that a new active site
can be generated through a single hydrophobic-to-ionizable amino acid
replacement that generates a partially buried group with perturbed
physico-chemical properties. We provide experimental and computational
evidence that conformational flexibility can assist the emergence and
subsequent evolution of new active sites by improving substrate and
transition-state binding, through the sampling of many potentially
productive conformations. Our results suggest a mechanism for the
emergence of primordial enzymes and highlight the potential of ancestral
reconstruction as a tool for protein engineering.