8SQW

particulate methane monooxygenase crosslinked with 2,2,2-trifluoroethanol bound

8SQW の概要

• エントリーDOI: 10.2210/pdb8sqw/pdb
• EMDBエントリー: 40714
• 分子名称: Particulate methane monooxygenase alpha subunit, Particulate methane monooxygenase beta subunit, Ammonia monooxygenase/methane monooxygenase, subunit C family protein, ... (10 entities in total)
• 機能のキーワード: metalloenzyme, inhibitor, membrane protein, nanodiscs, active site, oxidoreductase
• 由来する生物種: Methylococcus capsulatus; 詳細
• タンパク質・核酸の鎖数: 9
• 化学式量合計: 319249.51

構造登録者

Tucci, F.J.,Rosenzweig, A.C. (登録日: 2023-05-04, 公開日: 2023-11-15, 最終更新日: 2024-01-24)

主引用文献

Tucci, F.J.,Jodts, R.J.,Hoffman, B.M.,Rosenzweig, A.C.
Product analog binding identifies the copper active site of particulate methane monooxygenase.
Nat Catal, 6:1194-1204, 2023

PubMed Abstract: Nature's primary methane-oxidizing enzyme, the membrane-bound particulate methane monooxygenase (pMMO), catalyzes the oxidation of methane to methanol. pMMO activity requires copper, and decades of structural and spectroscopic studies have sought to identify the active site among three candidates: the Cu, Cu, and Cu sites. Challenges associated with the isolation of active pMMO have hindered progress toward locating its catalytic center. However, reconstituting pMMO into native lipid nanodiscs stabilizes its structure and recovers its activity. Here, these active samples were incubated with 2,2,2,-trifluoroethanol (TFE), a product analog that serves as a readily visualized active-site probe. Interactions of TFE with the Cu site were observed by both pulsed ENDOR spectroscopy and cryoEM, implicating Cu and the surrounding hydrophobic pocket as the likely site of methane oxidation. Use of these orthogonal techniques on parallel samples is a powerful approach that can circumvent difficulties in interpreting metalloenzyme cryoEM maps.

PubMed: 38187819
DOI: 10.1038/s41929-023-01051-x

実験手法

ELECTRON MICROSCOPY (2.16 Å)