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Related Experiment Video

Updated: Apr 10, 2026

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
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Native Top-Down Analysis of Membrane Protein Complexes Directly From In Vitro and Native Membranes.

Wonhyeuk Jung1, Aniruddha Panda1, Jaywon Lee1

  • 1Nanobiology Institute, Yale University, West Haven, Connecticut, USA; Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut, USA.

Molecular & Cellular Proteomics : MCP
|May 16, 2025
PubMed
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Native mass spectrometry now detects protein complexes directly from cell membranes. Gas phase supercharging enables analysis of intact protein-lipid complexes and their modifications in native vesicles.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Cell Biology

Background:

  • Macromolecular organization of proteins and lipids in cellular membranes is crucial for cell function.
  • Native mass spectrometry (nMS) is a powerful tool for studying these associations.
  • Current nMS methods often require protein extraction into detergent environments.

Purpose of the Study:

  • To extend gas phase supercharging methodology to native cell-derived membrane vesicles.
  • To develop a protocol for producing nMS-ready native membrane vesicles.
  • To demonstrate the detection and identification of protein complexes and proteoforms directly from native membranes.

Main Methods:

  • Supercharger-assisted pre-quadrupole activation followed by native top-down MS/MS.
Keywords:
drug bindingelectron capture fragmentationmembrane protein complexnative top–down mass spectrometryproteoform analysis

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Related Experiment Videos

Last Updated: Apr 10, 2026

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  • Combined collision-based and electron capture-based fragmentation approaches.
  • Development of a protocol for nMS-ready native membrane vesicle preparation.

    • Main Results:

    • Successfully detected and identified integral and membrane-associated protein complexes from E. coli total membranes.
    • Identified lipidated proteoforms of the hetero-pentameric BAM-complex and bound co-factors for DLDH.
    • Demonstrated the platform's utility for studying drug binding to membrane proteins, using BAM-complex as a model.

    Conclusions:

    • Supercharging-enabled native top-down mass spectrometry (nTD-platform) allows direct analysis of protein-lipid complexes in native membrane vesicles.
    • This approach preserves native protein associations and enables detailed proteoform and post-translational modification analysis.
    • The platform offers a novel method for drug discovery targeting membrane proteins.