Conformational switch triggered by α-ketoglutarate in a halogenase of curacin A biosynthesis

Abstract

The CurA halogenase (Hal) catalyzes a cryptic chlorination leading to cyclopropane ring formation in the synthesis of the natural product curacin A. Hal belongs to a family of enzymes that use Fe²⁺, O₂ and α-ketoglutarate (αKG) to perform a variety of halogenation reactions in natural product biosynthesis. Crystal structures of the enzyme in five ligand states reveal strikingly different open and closed conformations dependent on αKG binding. The open form represents ligand-free enzyme, preventing substrate from entering the active site until both αKG and chloride are bound, while the closed form represents the holoenzyme with αKG and chloride coordinated to iron. Candidate amino acid residues involved in substrate recognition were identified by site-directed mutagenesis. These new structures provide direct evidence of a conformational switch driven by αKG leading to chlorination of an early pathway intermediate.