4DXY
Crystal structures of CYP101D2 Y96A mutant
Summary for 4DXY
| Entry DOI | 10.2210/pdb4dxy/pdb |
| Related | 3NV5 |
| Descriptor | Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE, DI(HYDROXYETHYL)ETHER, ... (4 entities in total) |
| Functional Keywords | cytochrome p450 mutant, haem-dependent, mono-oxygenases, oxidoreductase |
| Biological source | Novosphingobium aromaticivorans |
| Total number of polymer chains | 1 |
| Total formula weight | 47946.42 |
| Authors | Zhou, W.,Bell, S.G.,Yang, W.,Dale, A.,Wong, L.-L. (deposition date: 2012-02-28, release date: 2012-08-15, Last modification date: 2023-11-08) |
| Primary citation | Bell, S.G.,Yang, W.,Dale, A.,Zhou, W.,Wong, L.-L. Improving the affinity and activity of CYP101D2 for hydrophobic substrates Appl.Microbiol.Biotechnol., 2012 Cited by PubMed Abstract: CYP101D2 is a cytochrome P450 monooxygenase from Novosphingobium aromaticivorans which is closely related to CYP101A1 (P450cam) from Pseudomonas putida. Both enzymes selectively hydroxylate camphor to 5-exo-hydroxycamphor, and the residues that line the active sites of both enzymes are similar including the pre-eminent Tyr96 residue. However, Met98 and Leu253 in CYP101D2 replace Phe98 and Val247 in CYP101A1, and camphor binding only results in a maximal change in the spin state to 40 % high-spin. Substitutions at Tyr96, Met98 and Leu253 in CYP101D2 reduced both the spin state shift on camphor binding and the camphor oxidation activity. The Tyr96Ala mutant increased the affinity of CYP101D2 for hydrocarbon substrates including adamantane, cyclooctane, hexane and 2-methylpentane. The monooxygenase activity of the Tyr96Ala variant towards alkane substrates was also enhanced compared with the wild-type enzyme. The crystal structure of the substrate-free form of this variant shows the enzyme in an open conformation (PDB: 4DXY), similar to that observed with the wild-type enzyme (PDB: 3NV5), with the side chain of Ala96 pointing away from the heme. Despite this, the binding and activity data suggest that this residue plays an important role in substrate binding, evidencing that the enzyme probably undergoes catalysis in a more closed conformation, similar to those observed in the crystal structures of CYP101A1 (PDB: 2CPP) and CYP101D1 (PDB: 3LXI). PubMed: 22820521DOI: 10.1007/s00253-012-4278-7 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
Download full validation report
