7S3T

NzeB Diketopiperazine Dimerase Mutant: Q68I-G87A-A89G-I90V

7S3T の概要

• エントリーDOI: 10.2210/pdb7s3t/pdb
• 分子名称: NascB, PROTOPORPHYRIN IX CONTAINING FE, (3S,8aS)-3-(1H-indol-3-ylmethyl)hexahydropyrrolo[1,2-a]pyrazine-1,4-dione, ... (6 entities in total)
• 機能のキーワード: cytochrome p450, diketopiperazine dimerase, natural products, biosynthetic protein
• 由来する生物種: Streptomyces sp.
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 45165.08

構造登録者

Harris, N.R.,Shende, V.V.,Sanders, J.N.,Newmister, S.A.,Khatri, Y.,Movassaghi, M.,Houk, K.N.,Sherman, D.H. (登録日: 2021-09-08, 公開日: 2022-10-05, 最終更新日: 2023-11-15)

主引用文献

Shende, V.V.,Harris, N.R.,Sanders, J.N.,Newmister, S.A.,Khatri, Y.,Movassaghi, M.,Houk, K.N.,Sherman, D.H.
Molecular Dynamics Simulations Guide Chimeragenesis and Engineered Control of Chemoselectivity in Diketopiperazine Dimerases.
Angew.Chem.Int.Ed.Engl., 2023

PubMed Abstract: In the biosynthesis of the tryptophan-linked dimeric diketopiperazines (DKPs), cytochromes P450 selectively couple DKP monomers to generate a variety of intricate and isomeric frameworks. To determine the molecular basis for selectivity of these biocatalysts we obtained a high-resolution crystal structure of selective Csp -N bond forming dimerase, AspB. Overlay of the AspB structure onto C-C and C-N bond forming homolog NzeB revealed no significant structural variance to explain their divergent chemoselectivities. Molecular dynamics (MD) simulations identified a region of NzeB with increased conformational flexibility relative to AspB, and interchange of this region along with a single active site mutation led to a variant that catalyzes exclusive C-N bond formation. MD simulations also suggest that intermolecular C-C or C-N bond formation results from a change in mechanism, supported experimentally through use of a substrate mimic.

PubMed: 36610039
DOI: 10.1002/anie.202210254

実験手法

X-RAY DIFFRACTION (1.4 Å)