7X7Z

The crystal structure of 2+2/4+2 cyclase PloI4

7X7Z の概要

• エントリーDOI: 10.2210/pdb7x7z/pdb
• 分子名称: PloI4 (2 entities in total)
• 機能のキーワード: ploi4, 2+2/4+2 cyclase, lyase
• 由来する生物種: Micromonospora sp.
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 61992.64

構造登録者

Li, M.,Pan, L.F. (登録日: 2022-03-10, 公開日: 2023-02-08, 最終更新日: 2023-11-29)

主引用文献

Wang, H.,Zou, Y.,Li, M.,Tang, Z.,Wang, J.,Tian, Z.,Strassner, N.,Yang, Q.,Zheng, Q.,Guo, Y.,Liu, W.,Pan, L.,Houk, K.N.
A cyclase that catalyses competing 2 + 2 and 4 + 2 cycloadditions.
Nat.Chem., 15:177-184, 2023

PubMed Abstract: Cycloaddition reactions are among the most widely used reactions in chemical synthesis. Nature achieves these cyclization reactions with a variety of enzymes, including Diels-Alderases that catalyse concerted 4 + 2 cycloadditions, but biosynthetic enzymes with 2 + 2 cyclase activity have yet to be discovered. Here we report that PloI4, a β-barrel-fold protein homologous to the exo-selective 4 + 2 cyclase that functions in the biosynthesis of pyrroindomycins, catalyses competitive 2 + 2 and 4 + 2 cycloaddition reactions. PloI4 is believed to catalyse an endo-4 + 2 cycloaddition in the biosynthesis of pyrrolosporin A; however, when the substrate precursor of pyrroindomycins was treated with PloI4, an exo-2 + 2 adduct was produced in addition to the exo- and endo-4 + 2 adducts. Biochemical characterizations, computational analyses, (co)crystal structures and mutagenesis outcomes have allowed the catalytic versatility of PloI4 to be rationalized. Mechanistic studies involved the directed engineering of PloI4 to variants that produced the exo-4 + 2, endo-4 + 2 or exo-2 + 2 product preferentially. This work illustrates an enzymatic thermal 2 + 2 cycloaddition and provides evidence of a process through which an enzyme evolves along with its substrate for specialization and activity improvement.

PubMed: 36690833
DOI: 10.1038/s41557-022-01104-x

実験手法

X-RAY DIFFRACTION (2.903 Å)