6CUN

Engineered Cytochrome c from Rhodothermus marinus (Rma TDE) bound to carbene intermediate (1-ethoxy-1-oxopropan-2-ylidene)

6CUN の概要

• エントリーDOI: 10.2210/pdb6cun/pdb
• 関連するPDBエントリー: 6CUK
• 分子名称: Cytochrome c, HEME C, ETHYL PROPIONATE, ... (4 entities in total)
• 機能のキーワード: carbene transferase, silicon-carbon bond, lyase
• 由来する生物種: Rhodothermus marinus (Rhodothermus obamensis)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14750.42

構造登録者

Lewis, R.D.,Buller, A.R.,Arnold, F.H. (登録日: 2018-03-26, 公開日: 2018-06-27, 最終更新日: 2024-11-13)

主引用文献

Lewis, R.D.,Garcia-Borras, M.,Chalkley, M.J.,Buller, A.R.,Houk, K.N.,Kan, S.B.J.,Arnold, F.H.
Catalytic iron-carbene intermediate revealed in a cytochromeccarbene transferase.
Proc. Natl. Acad. Sci. U.S.A., 115:7308-7313, 2018

PubMed Abstract: Recently, heme proteins have been discovered and engineered by directed evolution to catalyze chemical transformations that are biochemically unprecedented. Many of these nonnatural enzyme-catalyzed reactions are assumed to proceed through a catalytic iron porphyrin carbene (IPC) intermediate, although this intermediate has never been observed in a protein. Using crystallographic, spectroscopic, and computational methods, we have captured and studied a catalytic IPC intermediate in the active site of an enzyme derived from thermostable () cytochrome High-resolution crystal structures and computational methods reveal how directed evolution created an active site for carbene transfer in an electron transfer protein and how the laboratory-evolved enzyme achieves perfect carbene transfer stereoselectivity by holding the catalytic IPC in a single orientation. We also discovered that the IPC in cytochrome has a singlet ground electronic state and that the protein environment uses geometrical constraints and noncovalent interactions to influence different IPC electronic states. This information helps us to understand the impressive reactivity and selectivity of carbene transfer enzymes and offers insights that will guide and inspire future engineering efforts.

PubMed: 29946033
DOI: 10.1073/pnas.1807027115

実験手法

X-RAY DIFFRACTION (1.29 Å)