Engineered reactivity of a bacterial E1-like enzyme enables ATP-driven modification of protein C termini.

ABSTRACT: In biological systems, ATP provides an energetic driving force for peptide bond formation, but protein chemists lack tools that emulate this strategy. Inspired by the eukaryotic ubiquitination cascade, we developed an ATP-driven platform for C-terminal activation and peptide ligation based on E. coli MccB, a bacterial ancestor of ubiquitin-activating (E1) enzymes that natively catalyzes C-terminal phosphoramidate bond formation. We show that MccB can act on non-native substrates to generate an O-AMPylated electrophile that can react with exogenous nucleophiles to form diverse C-terminal functional groups including thioesters, a versatile class of biological intermediates that have been exploited for protein semisynthesis. To direct this activity towards specific proteins of i

SUBMITTER: Frazier CL 

PROVIDER: S-EPMC11118369 | biostudies-literature | 2024 May

REPOSITORIES: biostudies-literature