A Schiff base intermediate
in TYW1 is visualized. (A) An intact
[4Fe–4S] auxiliary cluster is shown surrounded by 2Fo–Fc
electron density, contoured at 1 σ (blue). Three cysteine ligands,
C26, C39, and C52, bind the auxiliary cluster, producing a site differentiated
iron ion, which is in close proximity to K41. A positive difference
density (green) contoured at +3 σ is shown extending from Nz
of K41 to the unique iron. (B) To account for difference electron
density, a Schiff base between K41 and the substrate pyruvate (KAC41)
was modeled and refined. A 2Fo–Fc omit density map (cyan) contoured
at 1 σ is shown for the auxiliary cluster, cysteine ligands,
and covalent adduct. (C) The Schiff base intermediate, KAC41, ligates
the unique iron of the auxiliary cluster of TYW1 in a bidentate fashion
through interactions with the Nz atom (blue) and a carboxylic oxygen
(red). (D) SAM ligating the unique iron of the SAM cluster in MoaA
(PDB 1TV8),
utilizing the amino nitrogen (blue) and a carboxylic oxygen (red).
All residues are shown as sticks.