@article{c9746fd31b264f31b566ba07dbd9a862,
title = "Chemical Additives Enable Native Mass Spectrometry Measurement of Membrane Protein Oligomeric State within Intact Nanodiscs",
abstract = "Membrane proteins play critical biochemical roles but remain challenging to study. Recently, native or nondenaturing mass spectrometry (MS) has made great strides in characterizing membrane protein interactions. However, conventional native MS relies on detergent micelles, which may disrupt natural interactions. Lipoprotein nanodiscs provide a platform to present membrane proteins for native MS within a lipid bilayer environment, but previous native MS of membrane proteins in nanodiscs has been limited by the intermediate stability of nanodiscs. It is difficult to eject membrane proteins from nanodiscs for native MS but also difficult to retain intact nanodisc complexes with membrane proteins inside. Here, we employed chemical reagents that modulate the charge acquired during electrospray ionization (ESI). By modulating ESI conditions, we could either eject the membrane protein complex with few bound lipids or capture the intact membrane protein nanodisc complex - allowing measurement of the membrane protein oligomeric state within an intact lipid bilayer environment. The dramatic differences in the stability of nanodiscs under different ESI conditions opens new applications for native MS of nanodiscs.",
author = "Keener, {James E.} and Zambrano, {Dane Evan} and Guozhi Zhang and Zak, {Ciara K.} and Reid, {Deseree J.} and Deodhar, {Bhushan S.} and Pemberton, {Jeanne E.} and Prell, {James S.} and Marty, {Michael T.}",
note = "Funding Information: The authors thank Maria Reinhardt-Szyba, Kyle Fort, and Alexander Makarov at Thermo Fisher Scientific for support on the ultrahigh mass range modification of the Q-Exactive HF instrument. The pMSP1D1 and pMSP1E3D1 plasmids were gifts from Stephen Sligar (Addgene plasmid nos. 20061 and 20066). The authors thank Wolfgang Peti for use of the nanoDSF instrumentation and Elaine Marzluff for helpful discussions. This work was funded by an American Cancer Society Institutional Research Grant (IRG-16-124-37-IRG), the Bisgrove Scholar Award from Science Foundation Arizona, the American Society for Mass Spectrometry Research Award, and the National Institute of General Medical Sciences and National Institutes of Health under Award Numbers R01 GM127579 and R35 GM128624. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. J.E.P. and B.S.D. gratefully acknowledge support of part of this research through an award from the National Science Foundation (CHE-1339597), jointly funded by the Environmental Protection Agency, as part of the Networks for Sustainable Molecular Design and Synthesis Program. J.S.P. thanks the National Institute of Allergy and Infectious Diseases for generous support (Grant No. R21-AI125804-02). Publisher Copyright: {\textcopyright} 2018 American Chemical Society.",
year = "2019",
month = jan,
day = "16",
doi = "10.1021/jacs.8b11529",
language = "English (US)",
volume = "141",
pages = "1054--1061",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "2",
}