8DSV
The structure of NicA2 in complex with N-methylmyosmine
Summary for 8DSV
| Entry DOI | 10.2210/pdb8dsv/pdb |
| Descriptor | FAD-dependent oxidoreductase, DIHYDROFLAVINE-ADENINE DINUCLEOTIDE, TETRAETHYLENE GLYCOL, ... (6 entities in total) |
| Functional Keywords | nicotine-degrading enzyme, flavoprotein, oxidoreductase |
| Biological source | Pseudomonas putida |
| Total number of polymer chains | 2 |
| Total formula weight | 97465.37 |
| Authors | Wu, K.,Dulchavsky, M.,Li, J.,Bardwell, J.C.A. (deposition date: 2022-07-22, release date: 2023-07-26, Last modification date: 2023-11-08) |
| Primary citation | Dulchavsky, M.,Mitra, R.,Wu, K.,Li, J.,Boer, K.,Liu, X.,Zhang, Z.,Vasquez, C.,Clark, C.T.,Funckes, K.,Shankar, K.,Bonnet-Zahedi, S.,Siddiq, M.,Sepulveda, Y.,Suhandynata, R.T.,Momper, J.D.,Calabrese, A.N.,George, O.,Stull, F.,Bardwell, J.C.A. Directed evolution unlocks oxygen reactivity for a nicotine-degrading flavoenzyme. Nat.Chem.Biol., 19:1406-1414, 2023 Cited by PubMed Abstract: The flavoenzyme nicotine oxidoreductase (NicA2) is a promising injectable treatment to aid in the cessation of smoking, a behavior responsible for one in ten deaths worldwide. NicA2 acts by degrading nicotine in the bloodstream before it reaches the brain. Clinical use of NicA2 is limited by its poor catalytic activity in the absence of its natural electron acceptor CycN. Without CycN, NicA2 is instead oxidized slowly by dioxygen (O), necessitating unfeasibly large doses in a therapeutic setting. Here, we report a genetic selection strategy that directly links CycN-independent activity of NicA2 to growth of Pseudomonas putida S16. This selection enabled us to evolve NicA2 variants with substantial improvement in their rate of oxidation by O. The encoded mutations cluster around a putative O tunnel, increasing flexibility and accessibility to O in this region. These mutations further confer desirable clinical properties. A variant form of NicA2 is tenfold more effective than the wild type at degrading nicotine in the bloodstream of rats. PubMed: 37770699DOI: 10.1038/s41589-023-01426-y PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.5 Å) |
Structure validation
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