Structure and activity of particulate methane monooxygenase arrays in methanotrophs.

Structure and activity of particulate methane monooxygenase arrays in methanotrophs.
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DOI:
10.1038/s41467-022-32752-9
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发表时间:
2022-09-05
影响因子:
16.6
通讯作者:
--
中科院分区:
综合性期刊1区
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--
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甲烷氧化细菌在温室气体减排中发挥着重要作用,在生物制造中具有潜在的应用前景。它们的主要代谢酶,颗粒甲烷单加氧酶(pMMO),被安置在铜诱导的胞内质膜(ICM),其功能和生物起源尚不清楚。我们表明,通过连续冷冻聚焦离子束(cryoFIB)铣削/扫描电子显微镜(SEM)体积成像和基于板层的细胞冷冻电子断层扫描(cryoET),这些ICM来自细胞内膜。pMMO三聚体,解决了冷冻ET和subtomography平均为4.8 μ m的ICM,形成更高阶的六边形阵列在完整的细胞。阵列形成与酶活性增加相关,突出了在其天然环境中研究酶的重要性。这些发现也证明了cryoET在细胞环境中对天然膜酶进行结构表征的能力。颗粒甲烷单加氧酶(pMMO)是甲烷氧化菌利用的主要酶。在这里,作者通过冷冻电子断层扫描确定了pMMO的天然结构,揭示了完整细胞中脂质稳定的特征和更高阶的六边形阵列排列。
Methane-oxidizing bacteria play a central role in greenhouse gas mitigation and have potential applications in biomanufacturing. Their primary metabolic enzyme, particulate methane monooxygenase (pMMO), is housed in copper-induced intracytoplasmic membranes (ICMs), of which the function and biogenesis are not known. We show by serial cryo-focused ion beam (cryoFIB) milling/scanning electron microscope (SEM) volume imaging and lamellae-based cellular cryo-electron tomography (cryoET) that these ICMs are derived from the inner cell membrane. The pMMO trimer, resolved by cryoET and subtomogram averaging to 4.8 Å in the ICM, forms higher-order hexagonal arrays in intact cells. Array formation correlates with increased enzymatic activity, highlighting the importance of studying the enzyme in its native environment. These findings also demonstrate the power of cryoET to structurally characterize native membrane enzymes in the cellular context. Particulate methane monooxygenase (pMMO) is the main enzyme used by methanotrophs. Here, the authors determined the native structure of pMMO by cryo-electron tomography, revealing lipid-stabilized features and a higher-order hexagonal array arrangement in intact cells.
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