5OA7
Fe(II)/(alpha)ketoglutarate-dependent dioxygenase AsqJ_V72I mutant in complex with cyclopeptin (1b)
Summary for 5OA7
| Entry DOI | 10.2210/pdb5oa7/pdb |
| Related | 5DAP |
| Descriptor | Iron/alpha-ketoglutarate-dependent dioxygenase asqJ, NICKEL (II) ION, 2-OXOGLUTARIC ACID, ... (5 entities in total) |
| Functional Keywords | antibiotics, quinolone biosynthesis, molecular engineering, desaturase, catalytic mechanism, mutagenesis, pi-stacking, oxidoreductase |
| Biological source | Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139) |
| Total number of polymer chains | 1 |
| Total formula weight | 34478.00 |
| Authors | Groll, M.,Braeuer, A.,Kaila, V.R.I. (deposition date: 2017-06-21, release date: 2018-04-04, Last modification date: 2024-01-17) |
| Primary citation | Mader, S.L.,Brauer, A.,Groll, M.,Kaila, V.R.I. Catalytic mechanism and molecular engineering of quinolone biosynthesis in dioxygenase AsqJ. Nat Commun, 9:1168-1168, 2018 Cited by PubMed Abstract: The recently discovered Fe/α-ketoglutarate-dependent dioxygenase AsqJ from Aspergillus nidulans stereoselectively catalyzes a multistep synthesis of quinolone alkaloids, natural products with significant biomedical applications. To probe molecular mechanisms of this elusive catalytic process, we combine here multi-scale quantum and classical molecular simulations with X-ray crystallography, and in vitro biochemical activity studies. We discover that methylation of the substrate is essential for the activity of AsqJ, establishing molecular strain that fine-tunes π-stacking interactions within the active site. To rationally engineer AsqJ for modified substrates, we amplify dispersive interactions within the active site. We demonstrate that the engineered enzyme has a drastically enhanced catalytic activity for non-methylated surrogates, confirming our computational data and resolved high-resolution X-ray structures at 1.55 Å resolution. Our combined findings provide crucial mechanistic understanding of the function of AsqJ and showcase how combination of computational and experimental data enables to rationally engineer enzymes. PubMed: 29563492DOI: 10.1038/s41467-018-03442-2 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.65 Å) |
Structure validation
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