7TC7

Cryo-EM structure of methane monooxygenase hydroxylase (by quantifoil)

7TC7 の概要

• エントリーDOI: 10.2210/pdb7tc7/pdb
• EMDBエントリー: 25804
• 分子名称: Methane monooxygenase component A alpha chain, Methane monooxygenase component A beta chain, Methane monooxygenase component A gamma chain, ... (4 entities in total)
• 機能のキーワード: methane monooxygenase, hydroxylase, cryo-em, electron transport, oxidoreductase
• 由来する生物種: Methylococcus capsulatus; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 251786.45

構造登録者

Cho, U.S.,Kim, B.C. (登録日: 2021-12-23, 公開日: 2023-01-25, 最終更新日: 2025-05-28)

主引用文献

Ahn, E.,Kim, B.,Park, S.,Erwin, A.L.,Sung, S.H.,Hovden, R.,Mosalaganti, S.,Cho, U.S.
Batch Production of High-Quality Graphene Grids for Cryo-EM: Cryo-EM Structure of Methylococcus capsulatus Soluble Methane Monooxygenase Hydroxylase.
Acs Nano, 17:6011-6022, 2023

PubMed Abstract: Cryogenic electron microscopy (cryo-EM) has become a widely used tool for determining the protein structure. Despite recent technical advances, sample preparation remains a major bottleneck for several reasons, including protein denaturation at the air-water interface, the presence of preferred orientations, nonuniform ice layers, etc. Graphene, a two-dimensional allotrope of carbon consisting of a single atomic layer, has recently gained attention as a near-ideal support film for cryo-EM that can overcome these challenges because of its superior properties, including mechanical strength and electrical conductivity. Here, we introduce a reliable, easily implemented, and reproducible method to produce 36 graphene-coated grids within 1.5 days. To demonstrate their practical application, we determined the cryo-EM structure of soluble methane monooxygenase hydroxylase (sMMOH) at resolutions of 2.9 and 2.5 Å using Quantifoil and graphene-coated grids, respectively. We found that the graphene-coated grid has several advantages, including a smaller amount of protein required and avoiding protein denaturation at the air-water interface. By comparing the cryo-EM structure of sMMOH with its crystal structure, we identified subtle yet significant geometrical changes at the nonheme diiron center, which may better indicate the active site configuration of sMMOH in the resting/oxidized state.

PubMed: 36926824
DOI: 10.1021/acsnano.3c00463

実験手法

ELECTRON MICROSCOPY (2.9 Å)