Axial ligation of Fe(II)-bleomycin probed by XANES spectroscopy.

Full multiple scattering calculations of the Fe K-edge X-ray absorption near edge structure of bleomycin have been performed. Structural insight is based on the comparison between experimental and theoretical data calculated for different active site models coming from NMR-informed molecular dynamic simulations. In all models considered, the equatorial ligands (secondary amine in beta-aminoalanine, pyrimidine and imidazole rings and the beta-hydroxyhistidine) were left unchanged. Seven models with two axial ligands (the primary amine in beta-aminoalanine and the carbomoyl group of the mannose or a solvent molecule) were tested. The best agreement between theoretical and experimental spectra is achieved for the model of bleomycin with the primary amine and the oxygen of the mannose sugar occupying the axial positions. The coordination environment is characterized by serious distortions of the Fe octahedron, including the presence of one ligand with a very short bond length and significant angular distortions.