4L4F
Structure of cyanide and camphor bound P450cam mutant L358A/K178G/D182N
Summary for 4L4F
| Entry DOI | 10.2210/pdb4l4f/pdb |
| Related | 4L49 4L4A 4L4B 4L4C 4L4D 4L4E 4L4G |
| Descriptor | Camphor 5-monooxygenase, PROTOPORPHYRIN IX CONTAINING FE, CYANIDE ION, ... (6 entities in total) |
| Functional Keywords | mono-oxygenase, cytochrome p450, cyanide complex, oxidoreductase |
| Biological source | Pseudomonas putida |
| Cellular location | Cytoplasm : P00183 |
| Total number of polymer chains | 1 |
| Total formula weight | 47445.75 |
| Authors | Batabyal, D.,Li, H.,Poulos, T.L. (deposition date: 2013-06-07, release date: 2013-07-31, Last modification date: 2023-09-20) |
| Primary citation | Batabyal, D.,Li, H.,Poulos, T.L. Synergistic Effects of Mutations in Cytochrome P450cam Designed To Mimic CYP101D1. Biochemistry, 52:5396-5402, 2013 Cited by PubMed Abstract: A close orthologue to cytochrome P450cam (CYP101A1) that catalyzes the same hydroxylation of camphor to 5-exo-hydroxycamphor is CYP101D1. There are potentially important differences in and around the active site that could contribute to subtle functional differences. Adjacent to the heme iron ligand, Cys357, is Leu358 in P450cam, whereas this residue is Ala in CYP101D1. Leu358 plays a role in binding of the P450cam redox partner, putidaredoxin (Pdx). On the opposite side of the heme, about 15-20 Å away, Asp251 in P450cam plays a critical role in a proton relay network required for O2 activation but forms strong ion pairs with Arg186 and Lys178. In CYP101D1 Gly replaces Lys178. Thus, the local electrostatic environment and ion pairing are substantially different in CYP101D1. These sites have been systematically mutated in P450cam to the corresponding residues in CYP101D1 and the mutants analyzed by crystallography, kinetics, and UV-vis spectroscopy. Individually, the mutants have little effect on activity or structure, but in combination there is a major drop in enzyme activity. This loss in activity is due to the mutants being locked in the low-spin state, which prevents electron transfer from the P450cam redox partner, Pdx. These studies illustrate the strong synergistic effects on well-separated parts of the structure in controlling the equilibrium between the open (low-spin) and closed (high-spin) conformational states. PubMed: 23865948DOI: 10.1021/bi400676d PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.294 Å) |
Structure validation
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