8DSV

The structure of NicA2 in complex with N-methylmyosmine

8DSV の概要

• エントリーDOI: 10.2210/pdb8dsv/pdb
• 分子名称: FAD-dependent oxidoreductase, DIHYDROFLAVINE-ADENINE DINUCLEOTIDE, TETRAETHYLENE GLYCOL, ... (6 entities in total)
• 機能のキーワード: nicotine-degrading enzyme, flavoprotein, oxidoreductase
• 由来する生物種: Pseudomonas putida
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 97465.37

構造登録者

Wu, K.,Dulchavsky, M.,Li, J.,Bardwell, J.C.A. (登録日: 2022-07-22, 公開日: 2023-07-26, 最終更新日: 2023-11-08)

主引用文献

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

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: 37770699
DOI: 10.1038/s41589-023-01426-y

実験手法

X-RAY DIFFRACTION (2.5 Å)