Computationally Guided Redesign of a Heme-free Cytochrome with Native-like Structure and Stability.

ABSTRACT: Metals can play key roles in stabilizing protein structures, but ensuring their proper incorporation is a challenge when a metalloprotein is overexpressed in a non-native cellular environment. Here, we have used computational protein design tools to redesign cytochrome b₅₆₂ (cyt b₅₆₂), which relies on the binding of its heme cofactor to achieve its proper fold, into a stable, heme-free protein. The resulting protein, ApoCyt, features only four mutations and no metal-ligand or covalent bonds, yet displays improved stability over cyt b₅₆₂. Mutagenesis studies and X-ray crystal structures reveal that the increase in stability is due to the computationally prescribed mutations, which stabilize the protein fold through a combination of hydro

SUBMITTER: Hoffnagle AM 

PROVIDER: S-EPMC9949987 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature