9CCR

Crystal structure of the EspE7 thioesterase mutant R35A from the esperamicin biosynthetic pathway at 1.6 A

9CCR の概要

• エントリーDOI: 10.2210/pdb9ccr/pdb
• 関連するPDBエントリー: 5vpj
• 分子名称: Thioesterase, POTASSIUM ION, DODECYL-COA, ... (4 entities in total)
• 機能のキーワード: thioesterase, hot-dog, enediyne biosynthetic pathway, esperamicin biosynthesis, natural product biosynthesis, hydrolase
• 由来する生物種: Actinomadura verrucosospora
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 84886.96

構造登録者

Miller, M.D.,Hankore, E.D.,Xu, W.,Kosgei, A.J.,Bhardwaj, M.,Thorson, J.S.,Van Lanen, S.G.,Phillips Jr., G.N. (登録日: 2024-06-23, 公開日: 2025-06-25, 最終更新日: 2025-11-05)

主引用文献

Hankore, E.D.,Miller, M.D.,Kosgei, A.J.,Xu, W.,Tan, K.,Endres, M.,Bhardwaj, M.,Joachimiak, G.,Joachimiak, A.,Phillips Jr., G.N.,Thorson, J.S.,Van Lanen, S.G.
Functional and Structural Studies on the Esperamicin Thioesterase and Progress toward Understanding Enediyne Core Biosynthesis.
J.Nat.Prod., 88:2360-2371, 2025

PubMed Abstract: Enediynes are among the most potent antitumor and antibacterial natural products. Studies on their biosynthetic pathways have identified a shared, linear polyene precursor generated from an iterative type I polyketide synthase (PKSE) as the source of the enediyne warhead. A key step is the release of this polyene from the PKSE by a discrete thioesterase (TE). Here, we used X-ray crystallography, site-directed mutagenesis, and heterologous coexpression of PKSEs and TEs to elucidate how enediyne TEs mediate the production of the polyene. We solved the structure of wild-type EspE7 from esperamicin producer . The substrate binding pocket was also defined upon serendipitous cocrystallization of an EspE7 mutant with a fatty acyl-CoA ligand. Structural data and activity assays with EspE7 mutants provide strong evidence that Glu68 in EspE7 and the analogous Glu residue in other enediyne TEs functions as a key catalytic residue, thus supporting a hydrolysis mechanism for enediyne TEs that aligns with that of sp. 4-HB-CoA TE. Furthermore, combinations of 9- and 10-membered enediyne PKSEs and TEs produced 1,3,5,7,9,11,13-pentadecaheptaene () as the major product. Thus, the data further support previous conclusions that serves as the sole precursor for the biosynthesis of all enediyne cores.

PubMed: 41066299
DOI: 10.1021/acs.jnatprod.5c00660

実験手法

X-RAY DIFFRACTION (1.57 Å)