3ZK5
PikC D50N mutant bound to the 10-DML analog with the 3-(N,N-dimethylamino)ethanoate anchoring group
Summary for 3ZK5
| Entry DOI | 10.2210/pdb3zk5/pdb |
| Related | 4B7S |
| Descriptor | CYTOCHROME P450 HYDROXYLASE PIKC, PROTOPORPHYRIN IX CONTAINING FE, (3R,4S,5S,7R,9E,11R,12R)-12-ethyl-3,5,7,11-tetramethyl-2,8-dioxooxacyclododec-9-en-4-yl N,N-dimethylglycinate, ... (4 entities in total) |
| Functional Keywords | oxidoreductase, monooxygenase, pikromycin biosynthesis |
| Biological source | STREPTOMYCES VENEZUELAE |
| Total number of polymer chains | 2 |
| Total formula weight | 98531.50 |
| Authors | Podust, L.M. (deposition date: 2013-01-21, release date: 2014-01-29, Last modification date: 2023-12-20) |
| Primary citation | Negretti, S.,Narayan, A.R.H.,Chiou, K.C.,Kells, P.M.,Stachowski, J.L.,Hansen, D.A.,Podust, L.M.,Montgomery, J.,Sherman, D.H. Directing Group-Controlled Regioselectivity in an Enzymatic C-H Bond Oxygenation. J.Am.Chem.Soc., 136:4901-, 2014 Cited by PubMed Abstract: Highly regioselective remote hydroxylation of a natural product scaffold is demonstrated by exploiting the anchoring mechanism of the biosynthetic P450 monooxygenase PikCD50N-RhFRED. Previous studies have revealed structural and biochemical evidence for the role of a salt bridge between the desosamine N,N-dimethylamino functionality of the natural substrate YC-17 and carboxylate residues within the active site of the enzyme, and selectivity in subsequent C-H bond functionalization. In the present study, a substrate-engineering approach was conducted that involves replacing desosamine with varied synthetic N,N-dimethylamino anchoring groups. We then determined their ability to mediate enzymatic total turnover numbers approaching or exceeding that of the natural sugar, while enabling ready introduction and removal of these amino anchoring groups from the substrate. The data establish that the size, stereochemistry, and rigidity of the anchoring group influence the regioselectivity of enzymatic hydroxylation. The natural anchoring group desosamine affords a 1:1 mixture of regioisomers, while synthetic anchors shift YC-17 analogue C-10/C-12 hydroxylation from 20:1 to 1:4. The work demonstrates the utility of substrate engineering as an orthogonal approach to protein engineering for modulation of regioselective C-H functionalization in biocatalysis. PubMed: 24627965DOI: 10.1021/JA5016052 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.89 Å) |
Structure validation
Download full validation report
