Inside-Out Design of Zinc-Binding Proteins with Non-Native Backbones.

ABSTRACT: The de novo design of functional proteins requires specification of tertiary structure and incorporation of molecular binding sites. Here, we develop an inside-out design strategy in the molecular modeling program Rosetta that begins with amino acid side chains from one or two α-helices making well-defined contacts with a ligand. A full-sized protein is then built around the ligand by adding additional helices that promote the formation of a protein core and allow additional contacts with the ligand. The protocol was tested by designing 12 zinc-binding proteins, each with 4-5 helices. Four of the designs were folded and bound to zinc with equilibrium dissociation constants varying between 95 nM and 1.1 μM. The design with the tightest affinity for zinc, N12, adopts a unique conforma

SUBMITTER: Guffy SL 

PROVIDER: S-EPMC9939277 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature