@article{4a96dd1041aa4707a0b81411426030d0,
title = "Native top-down mass spectrometry provides insights into the copper centers of membrane-bound methane monooxygenase",
abstract = "Aerobic methane oxidation is catalyzed by particulate methane monooxygenase (pMMO), a copper-dependent, membrane metalloenzyme composed of subunits PmoA, PmoB, and PmoC. Characterization of the copper active site has been limited by challenges in spectroscopic analysis stemming from the presence of multiple copper binding sites, effects of detergent solubilization on activity and crystal structures, and the lack of a heterologous expression system. Here we utilize nanodiscs coupled with native top-down mass spectrometry (nTDMS) to determine the copper stoichiometry in each pMMO subunit and to detect post-translational modifications (PTMs). These results indicate the presence of a mononuclear copper center in both PmoB and PmoC. pMMO-nanodisc complexes with a higher stoichiometry of copper-bound PmoC exhibit increased activity, suggesting that the PmoC copper site plays a role in methane oxidation activity. These results provide key insights into the pMMO copper centers and demonstrate the ability of nTDMS to characterize complex membrane-bound metalloenzymes.",
author = "Ro, {Soo Y.} and Schachner, {Luis F.} and Koo, {Christopher W.} and Rahul Purohit and Remis, {Jonathan P.} and Kenney, {Grace E.} and Liauw, {Brandon W.} and Thomas, {Paul M.} and Patrie, {Steven M.} and Kelleher, {Neil L.} and Rosenzweig, {Amy C.}",
note = "Funding Information: This work was funded by National Institutes of Health grants GM118035 (A.C.R.) and 1S10OD025194–01 (N.L.K). Research in this publication is also supported by a fellowship associated with the Chemistry of Life Processes Predoctoral Training Grant T32GM105538 at Northwestern University. L.F.S. is a Gilliam Fellow of the Howard Hughes Medical Institute. The Quantitative Bio-element Imaging Center at North-western is supported by NASA Ames Research Center NNA06CB93G. The Proteomics Center for Excellence and at Northwestern is supported by NIH grant NCI CCSG P30 CA060553. The support from the R.H. Lurie Comprehensive Cancer Center of North-western University to the Northwestern University Structural Biology Facility is acknowledged. The Gatan K2 direct electron detector was purchased with funds provided by the Chicago Biomedical Consortium with support from the Searle Funds at The Chicago Community Trust. Collection of electron microscopy data was supported by Chicago Biomedical Consortium Postdoctoral Research Award PDR-097 (R. P.) with support from the Searle Funds at The Chicago Community Trust. Publisher Copyright: {\textcopyright} 2019, The Author(s).",
year = "2019",
month = dec,
day = "1",
doi = "10.1038/s41467-019-10590-6",
language = "English (US)",
volume = "10",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}