Engineering Nanodisc Scaffold Proteins for Native Mass Spectrometry

Deseree J. Reid, James E. Keener, Andrew P. Wheeler, Dane Evan Zambrano, Jessica M. Diesing, Maria Reinhardt-Szyba, Alexander Makarov, Michael T. Marty

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

Lipoprotein nanodiscs are ideally suited for native mass spectrometry because they provide a relatively monodisperse nanoscale lipid bilayer environment for delivering membrane proteins into the gas phase. However, native mass spectrometry of nanodiscs produces complex spectra that can be challenging to assign unambiguously. To simplify interpretation of nanodisc spectra, we engineered a series of mutant membrane scaffold proteins (MSP) that do not affect nanodisc formation but shift the masses of nanodiscs in a controllable way, eliminating isobaric interference from the lipids. Moreover, by mixing two different belts before assembly, the stoichiometry of MSP is encoded in the peak shape, which allows the stoichiometry to be assigned unambiguously from a single spectrum. Finally, we demonstrate the use of mixed belt nanodiscs with embedded membrane proteins to confirm the dissociation of MSP prior to desolvation.

Original languageEnglish (US)
Pages (from-to)11189-11192
Number of pages4
JournalAnalytical Chemistry
Volume89
Issue number21
DOIs
StatePublished - Nov 7 2017
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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