Recovery of particulate methane monooxygenase structure and activity in a lipid bilayer

Christopher W. Koo, Frank J. Tucci, Yuan He, Amy C. Rosenzweig*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

75 Scopus citations

Abstract

Bacterial methane oxidation using the enzyme particulate methane monooxygenase (pMMO) contributes to the removal of environmental methane, a potent greenhouse gas. Crystal structures determined using inactive, detergent-solubilized pMMO lack several conserved regions neighboring the proposed active site. We show that reconstituting pMMO in nanodiscs with lipids extracted from the native organism restores methane oxidation activity. Multiple nanodisc-embedded pMMO structures determined by cryo–electron microscopy to 2.14- to 2.46-angstrom resolution reveal the structure of pMMO in a lipid environment. The resulting model includes stabilizing lipids, regions of the PmoA and PmoC subunits not observed in prior structures, and a previously undetected copper-binding site in the PmoC subunit with an adjacent hydrophobic cavity. These structures provide a revised framework for understanding and engineering pMMO function.

Original languageEnglish (US)
Pages (from-to)1287-1291
Number of pages5
JournalScience
Volume375
Issue number6586
DOIs
StatePublished - Mar 18 2022

Funding

This work was supported by National Institutes of Health grants R35GM118035 (A.C.R.), T32GM008382 (C.W.K.), T32GM105538 (F.J.T.), and R01GM135651 (Y.H.). This work used resources of the Northwestern University Structural Biology Facility, which is generously supported by the NCI CCSG P30 CA060553 grant awarded to the Robert H. Lurie Comprehensive Cancer Center. We acknowledge the use of the Ametek K3 direct electron detector, which was generously provided by R. A. Lamb (HHMI investigator). A portion of this research was supported by NIH grant U24GM129547 and performed at the PNCC at OHSU and accessed through EMSL (grid.436923.9), a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research. Some of this work was performed at the Stanford-SLAC Cryo-EM Center (S2C2) supported by the NIH Common Fund Transformative High-Resolution Cryoelectron Microscopy program (U24 GM129541). Some of this work was performed at the National Center for CryoEM Access and Training (NCCAT) and the Simons Electron Microscopy Center located at the New York Structural Biology Center, supported by the NIH Common Fund Transformative High-Resolution Cryoelectron Microscopy program (U24 GM129539) and by grants from the Simons Foundation (SF349247) and the NY State Assembly.

ASJC Scopus subject areas

  • General

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