7V6H

Crystal Structure of the SpnL

7V6H の概要

• エントリーDOI: 10.2210/pdb7v6h/pdb
• 分子名称: Cyclopropane fatty-acyl-phospholipid synthase-like methyltransferase, S-ADENOSYL-L-HOMOCYSTEINE (2 entities in total)
• 機能のキーワード: spnl, rauhut-currier reaction, spinosyn a, saccharopolyspora spinosa, biosynthetic protein, transferase
• 由来する生物種: Saccharopolyspora spinosa
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 62268.15

構造登録者

Wu, H.-H.,Ko, T.-P.,Liu, H.-W.,Tsai, M.-D. (登録日: 2021-08-20, 公開日: 2021-12-29, 最終更新日: 2024-05-29)

主引用文献

Choi, S.H.,Jeon, B.,Kim, N.,Wu, H.H.,Ko, T.P.,Ruszczycky, M.W.,Isiorho, E.A.,Liu, Y.N.,Keatinge-Clay, A.T.,Tsai, M.D.,Liu, H.W.
Evidence for an Enzyme-Catalyzed Rauhut-Currier Reaction during the Biosynthesis of Spinosyn A.
J.Am.Chem.Soc., 143:20291-20295, 2021

PubMed Abstract: The catalog of enzymes known to catalyze the nucleophile-assisted formation of C-C bonds is extremely small, and there is presently no definitive example of a biological Rauhut-Currier reaction. Biosynthesis of the polyketide insecticide spinosyn A in involves a [4 + 2]-cycloaddition and a subsequent intramolecular C-C bond formation catalyzed by SpnF and SpnL, respectively. Isotope tracer experiments and kinetic isotope effects, however, imply that the SpnL-catalyzed reaction proceeds without initial deprotonation of the substrate. The crystal structure of SpnL exhibits high similarity to SAM-dependent methyltransferases as well as SpnF. The residue Cys60 is also shown to reside in the SpnL active site, and the Cys60Ala SpnL mutant is found to be devoid of activity. Moreover, SpnL is covalently modified at Cys60 and irreversibly inactivated when it is coincubated with a fluorinated substrate analogue designed as a suicide inactivator of nucleophile-assisted C-C bond formation. These results suggest that SpnL catalyzes a biological Rauhut-Currier reaction.

PubMed: 34813308
DOI: 10.1021/jacs.1c09482

実験手法

X-RAY DIFFRACTION (3.054 Å)