8FIE
Crystal Structure of Erwinia tracheiphila CYP114 mutant - A261D
Summary for 8FIE
| Entry DOI | 10.2210/pdb8fie/pdb |
| Descriptor | Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total) |
| Functional Keywords | cytochrome p450, gibberellin, oxidoreductase |
| Biological source | Erwinia tracheiphila PSU-1 |
| Total number of polymer chains | 1 |
| Total formula weight | 49541.78 |
| Authors | Stewart Jr., C.E.,Alexander, L.E.,Nagel, R. (deposition date: 2022-12-16, release date: 2023-07-05, Last modification date: 2023-10-25) |
| Primary citation | Nagel, R.,Alexander, L.E.,Stewart Jr., C.E.,Peters, R.J. Dual factors required for cytochrome-P450-mediated hydrocarbon ring contraction in bacterial gibberellin phytohormone biosynthesis. Proc.Natl.Acad.Sci.USA, 120:e2221549120-e2221549120, 2023 Cited by PubMed Abstract: Cytochromes P450 (CYPs) are heme-thiolate monooxygenases that prototypically catalyze the insertion of oxygen into unactivated C-H bonds but are capable of mediating more complex reactions. One of the most remarked-upon alternative reactions occurs during biosynthesis of the gibberellin A (GA) phytohormones, involving hydrocarbon ring contraction with coupled aldehyde extrusion of -kaurenoic acid to form the first gibberellin intermediate. While the unusual nature of this reaction has long been noted, its mechanistic basis has remained opaque. Building on identification of the relevant CYP114 from bacterial GA biosynthesis, detailed structure-function studies are reported here, including development of in vitro assays as well as crystallographic analyses both in the absence and presence of substrate. These structures provided insight into enzymatic catalysis of this unusual reaction, as exemplified by identification of a key role for the "missing" acid from an otherwise highly conserved acid-alcohol pair of residues. Notably, the results demonstrate that ring contraction requires dual factors, both the use of a dedicated ferredoxin and absence of the otherwise conserved acidic residue, with exclusion of either limiting turnover to just the initiating and more straightforward hydroxylation. The results provide detailed insight into the enzymatic structure-function relationships underlying this fascinating reaction and support the use of a semipinacol mechanism for the unusual ring contraction reaction. PubMed: 37339230DOI: 10.1073/pnas.2221549120 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.26 Å) |
Structure validation
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