9U2X
Crystal structure of N244L variant from Bacillus subtilis
Summary for 9U2X
| Entry DOI | 10.2210/pdb9u2x/pdb |
| Descriptor | Deferrochelatase, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total) |
| Functional Keywords | bacillus subtilis, dyp, hydrogen peroxide, ros, oxidoreductase |
| Biological source | Bacillus subtilis |
| Total number of polymer chains | 6 |
| Total formula weight | 242066.45 |
| Authors | |
| Primary citation | Zitare, U.A.,Castro, M.A.,Scocozza, M.F.,Borges, P.T.,Martins, L.O.,Murgida, D.H. A Single Amino Acid Substitution Reprograms ROS Selectivity and Catalytic Function in DyP Peroxidases. Inorg.Chem., 2026 Cited by PubMed Abstract: Dye-decolorizing peroxidases (DyPs) are heme enzymes with broad substrate scope and increasing biotechnological relevance, yet the structural determinants governing their activation by reactive oxygen species (ROS) remain incompletely understood. Here, we identify dynamic ROS gating as a mechanism controlling activation and catalytic efficiency through a comparative study of two class I DyPs from (DyP) and (DyP), together with the DyP N244L variant. By combining steady-state kinetics, ROS-selective electroreductive activation assays, UV-vis and resonance Raman spectroscopy, X-ray crystallography, and molecular dynamics simulations, we establish a direct structure-function relationship linking distal heme pocket organization and access-tunnel architecture to enzyme-specific ROS preferences. DyP WT is preferentially activated via OH, whereas DyP relies mainly on HO. Remarkably, the single N244L substitution shifts the catalytic, structural, and dynamical properties of DyP toward those of DyP, making both enzymes nearly indistinguishable in ROS usage and catalytic efficiency. This convergence arises from a reorganization of distal hydrogen-bonding networks and loop-mediated reshaping of the access tunnels, which together bias ROS accessibility and lower the barrier for compound I formation. These findings establish ROS gating as the principal determinant of DyP activation and provide a mechanistic framework for tuning ROS selectivity through protein engineering. PubMed: 42295056DOI: 10.1021/acs.inorgchem.6c02237 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.67 Å) |
Structure validation
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