Standard Proteoforms and Their Complexes for Native Mass Spectrometry

Luis F. Schachner, Ashley N. Ives, John P. McGee, Rafael D. Melani, Jared O. Kafader, Philip D. Compton, Steven M. Patrie, Neil L. Kelleher*

*Corresponding author for this work

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Native mass spectrometry (nMS) is a technique growing at the interface of analytical chemistry, structural biology, and proteomics that enables the detection and partial characterization of non-covalent protein assemblies. Currently, the standardization and dissemination of nMS is hampered by technical challenges associated with instrument operation, benchmarking, and optimization over time. Here, we provide a standard operating procedure for acquiring high-quality native mass spectra of 30–300 kDa proteins using an Orbitrap mass spectrometer. By describing reproducible sample preparation, loading, ionization, and nMS analysis, we forward two proteoforms and three complexes as possible standards to advance training and longitudinal assessment of instrument performance. Spectral data for five standards can guide assessment of instrument parameters, data production, and data analysis. By introducing this set of standards and protocols, we aim to help normalize native mass spectrometry practices across labs and provide benchmarks for reproducibility and high-quality data production in the years ahead.

Original languageEnglish (US)
Pages (from-to)1190-1198
Number of pages9
JournalJournal of the American Society for Mass Spectrometry
Volume30
Issue number7
DOIs
StatePublished - Jul 15 2019

Keywords

  • Multi-proteoform complexes
  • Native mass spectrometry
  • Native top-down mass spectrometry
  • Proteoforms
  • Rigor and reproducibility
  • Standards

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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