6VY9

Crystal structure of NotF prenyltransferase

6VY9 の概要

• エントリーDOI: 10.2210/pdb6vy9/pdb
• 分子名称: Deoxybrevianamide E synthase notF (1 entity in total)
• 機能のキーワード: fungal indole prenyltransferase, transferase
• 由来する生物種: Aspergillus sp.
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 430657.69

構造登録者

Dan, Q.,Smith, J.L. (登録日: 2020-02-25, 公開日: 2021-02-03, 最終更新日: 2023-10-25)

主引用文献

Kelly, S.P.,Shende, V.V.,Flynn, A.R.,Dan, Q.,Ye, Y.,Smith, J.L.,Tsukamoto, S.,Sigman, M.S.,Sherman, D.H.
Data Science-Driven Analysis of Substrate-Permissive Diketopiperazine Reverse Prenyltransferase NotF: Applications in Protein Engineering and Cascade Biocatalytic Synthesis of (-)-Eurotiumin A.
J.Am.Chem.Soc., 144:19326-19336, 2022

PubMed Abstract: Prenyltransfer is an early-stage carbon-hydrogen bond (C-H) functionalization prevalent in the biosynthesis of a diverse array of biologically active bacterial, fungal, plant, and metazoan diketopiperazine (DKP) alkaloids. Toward the development of a unified strategy for biocatalytic construction of prenylated DKP indole alkaloids, we sought to identify and characterize a substrate-permissive C2 reverse prenyltransferase (PT). As the first tailoring event within the biosynthesis of cytotoxic notoamide metabolites, PT NotF catalyzes C2 reverse prenyltransfer of brevianamide F. Solving a crystal structure of NotF (in complex with native substrate and prenyl donor mimic dimethylallyl S-thiolodiphosphate (DMSPP)) revealed a large, solvent-exposed active site, intimating NotF may possess a significantly broad substrate scope. To assess the substrate selectivity of NotF, we synthesized a panel of 30 sterically and electronically differentiated tryptophanyl DKPs, the majority of which were selectively prenylated by NotF in synthetically useful conversions (2 to >99%). Quantitative representation of this substrate library and development of a descriptive statistical model provided insight into the molecular origins of NotF's substrate promiscuity. This approach enabled the identification of key substrate descriptors (electrophilicity, size, and flexibility) that govern the rate of NotF-catalyzed prenyltransfer, and the development of an "induced fit docking (IFD)-guided" engineering strategy for improved turnover of our largest substrates. We further demonstrated the utility of NotF in tandem with oxidative cyclization using flavin monooxygenase, BvnB. This one-pot, biocatalytic cascade enabled the first chemoenzymatic synthesis of the marine fungal natural product, (-)-eurotiumin A, in three steps and 60% overall yield.

PubMed: 36223664
DOI: 10.1021/jacs.2c06631

実験手法

X-RAY DIFFRACTION (3.19 Å)