Characterization of porphobilinogen deaminase mutants reveals that arginine-173 is crucial for polypyrrole elongation mechanism.

ABSTRACT: Porphobilinogen deaminase (PBGD), the third enzyme in the heme biosynthesis, catalyzes the sequential coupling of four porphobilinogen (PBG) molecules into a heme precursor. Mutations in PBGD are associated with acute intermittent porphyria (AIP), a rare metabolic disorder. We used Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) to demonstrate that wild-type PBGD and AIP-associated mutant R167W both existed as holoenzymes (E_holo) covalently attached to the dipyrromethane cofactor, and three intermediate complexes, ES, ES₂, and ES₃, where S represents PBG. In contrast, only ES₂ was detected in AIP-associated mutant R173W, indicating that the formation of ES₃ is inhibited. The R173W crystal structure in the ES₂-state revealed major rearrangements of the loops around the active site, compared to wild-type PBGD in the E_holo-state. These results contribute to elucidating the structural pathogenesis of two common AIP-associated mutations and reveal the important structural role of Arg173 in the polypyrrole elongation mechanism.

SUBMITTER: Bustad HJ 

PROVIDER: S-EPMC7907807 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature