A Strategy for Dissecting the Architectures of Native Macromolecular Assemblies

Nat Methods. 2015 Dec;12(12):1135-8. doi: 10.1038/nmeth.3617. Epub 2015 Oct 5.

Abstract

It remains particularly problematic to define the structures of native macromolecular assemblies, which are often of low abundance. Here we present a strategy for isolating complexes at endogenous levels from GFP-tagged transgenic cell lines. Using cross-linking mass spectrometry, we extracted distance restraints that allowed us to model the complexes' molecular architectures.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cross-Linking Reagents / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Exosomes / chemistry
  • Green Fluorescent Proteins / genetics
  • Liver / chemistry
  • Mass Spectrometry / methods*
  • Mice, Transgenic
  • Models, Molecular*
  • Multiprotein Complexes / chemistry*
  • Multiprotein Complexes / genetics
  • Protein Conformation*
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae Proteins / chemistry
  • Sensitivity and Specificity

Substances

  • Cross-Linking Reagents
  • Multiprotein Complexes
  • Saccharomyces cerevisiae Proteins
  • Green Fluorescent Proteins