Top-down characterization of endogenous protein complexes with native proteomics

Nat Chem Biol. 2018 Jan;14(1):36-41. doi: 10.1038/nchembio.2515. Epub 2017 Nov 13.

Abstract

Protein complexes exhibit great diversity in protein membership, post-translational modifications and noncovalent cofactors, enabling them to function as the actuators of many important biological processes. The exposition of these molecular features using current methods lacks either throughput or molecular specificity, ultimately limiting the use of protein complexes as direct analytical targets in a wide range of applications. Here, we apply native proteomics, enabled by a multistage tandem MS approach, to characterize 125 intact endogenous complexes and 217 distinct proteoforms derived from mouse heart and human cancer cell lines in discovery mode. The native conditions preserved soluble protein-protein interactions, high-stoichiometry noncovalent cofactors, covalent modifications to cysteines, and, remarkably, superoxide ligands bound to the metal cofactor of superoxide dismutase 2. These data enable precise compositional analysis of protein complexes as they exist in the cell and demonstrate a new approach that uses MS as a bridge to structural biology.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Humans
  • Mice
  • Multiprotein Complexes / chemistry*
  • Multiprotein Complexes / genetics
  • Protein Conformation
  • Protein Multimerization*
  • Protein Processing, Post-Translational
  • Protein Subunits / chemistry
  • Protein Subunits / genetics
  • Proteomics / methods*
  • Tandem Mass Spectrometry / methods*

Substances

  • Multiprotein Complexes
  • Protein Subunits