9U53
Crystal structure of the CYP154C2 Q230A from Streptomyces avermitilis
Summary for 9U53
| Entry DOI | 10.2210/pdb9u53/pdb |
| Descriptor | Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE, DI(HYDROXYETHYL)ETHER, ... (5 entities in total) |
| Functional Keywords | cytochrome p450, hydroxylase, androstenedione, oxidoreductase |
| Biological source | Streptomyces avermitilis |
| Total number of polymer chains | 1 |
| Total formula weight | 46168.23 |
| Authors | Xu, L.H. (deposition date: 2025-03-20, release date: 2025-09-24, Last modification date: 2026-04-08) |
| Primary citation | Yang, J.,Huang, J.,He, X.,Fushinobu, S.,Xu, L.H. Conformational gating in CYP154C2: Gln230-mediated substrate recognition and catalytic switching revealed by structural dynamics. Biosci.Biotechnol.Biochem., 89:1144-1153, 2025 Cited by PubMed Abstract: Previously, we reported that CYP154C2 from Streptomyces avermitilis is capable of catalyzing the 2α-hydroxylation of the two model substrates, testosterone (TES) and androstenedione (ASD), and resolved the closed structures of both the substrate-free form and the TES-bound form. In this study, we extend these findings by determining the open-conformation structures of the substrate-free and ASD-bound forms-a rare achievement among bacterial P450s. Structural analyses revealed coordinated conformational shifts in the FG helices, HI helices, and BC loop during open-to-closed transitions. Despite divergent overall conformations, both substrates positioned their C2 atoms near the heme iron, aligning for 2α-hydroxylation. Mutagenesis studies established Gln230's pivotal role in substrate recognition and catalytic activation. High-resolution crystallography (1.97 Å) of the Q230A mutant revealed polyethylene glycol-occupied catalytic pockets (indicating complete loss of TES binding) while maintaining the open conformation. These results provide atomic-level evidence that Gln230 coordinates both substrate-driven conformational gating and catalytic site optimization. PubMed: 40408304DOI: 10.1093/bbb/zbaf076 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.97 Å) |
Structure validation
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