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

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

doi:10.1126/science.abm3282

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

Molecular Biosciences

Research output: Contribution to journal › Article › peer-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 language	English (US)
Pages (from-to)	1287-1291
Number of pages	5
Journal	Science
Volume	375
Issue number	6586
DOIs	https://doi.org/10.1126/science.abm3282
State	Published - 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

Access to Document

10.1126/science.abm3282

Cite this

@article{3ebaa302043143a094cb88fd6a8317af,

title = "Recovery of particulate methane monooxygenase structure and activity in a lipid bilayer",

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.",

author = "Koo, {Christopher W.} and Tucci, {Frank J.} and Yuan He and Rosenzweig, {Amy C.}",

year = "2022",

month = mar,

day = "18",

doi = "10.1126/science.abm3282",

language = "English (US)",

volume = "375",

pages = "1287--1291",

journal = "Science",

issn = "0036-8075",

publisher = "American Association for the Advancement of Science",

number = "6586",

}

TY - JOUR

T1 - Recovery of particulate methane monooxygenase structure and activity in a lipid bilayer

AU - Koo, Christopher W.

AU - Tucci, Frank J.

AU - He, Yuan

AU - Rosenzweig, Amy C.

PY - 2022/3/18

Y1 - 2022/3/18

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85126646883&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85126646883&partnerID=8YFLogxK

U2 - 10.1126/science.abm3282

DO - 10.1126/science.abm3282

M3 - Article

C2 - 35298269

AN - SCOPUS:85126646883

SN - 0036-8075

VL - 375

SP - 1287

EP - 1291

JO - Science

JF - Science

IS - 6586

ER -

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

Abstract

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

CryoEM structure of Methylocystis sp. str. Rockwell pMMO in a POPC nanodisc at 2.42 Angstrom resolution

CryoEM structure of Methylococcus capsulatus (Bath) pMMO treated with potassium cyanide in a native lipid nanodisc at 3.65 Angstrom resolution

CryoEM structure of Methylococcus capsulatus (Bath) pMMO in a native lipid nanodisc at 2.26 Angstrom resolution

Cite this

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

Abstract

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Datasets

CryoEM structure of Methylocystis sp. str. Rockwell pMMO in a POPC nanodisc at 2.42 Angstrom resolution

CryoEM structure of Methylococcus capsulatus (Bath) pMMO treated with potassium cyanide in a native lipid nanodisc at 3.65 Angstrom resolution

CryoEM structure of Methylococcus capsulatus (Bath) pMMO in a native lipid nanodisc at 2.26 Angstrom resolution

Cite this