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Identification and Regulation of Multimeric Protein Complexes in Autophagy via SILAC-Based Mass Spectrometry Approaches.
Journal article

Identification and Regulation of Multimeric Protein Complexes in Autophagy via SILAC-Based Mass Spectrometry Approaches.

  • Kaeser-Pebernard S Department of Biology, University of Fribourg, Fribourg, Switzerland.
  • Diedrich B Agilent Technologies, Waldbronn, Germany.
  • Dengjel J Department of Biology, University of Fribourg, Fribourg, Switzerland. joern.dengjel@unifr.ch.
  • 2019-01-06
Published in:
  • Methods in molecular biology (Clifton, N.J.). - 2019
AP-MS
Autophagy
Macroautophagy
Mass spectrometry
Proteomics
SEC-PCP-SILAC
SILAC
Starvation
Autophagy
Chromatography, Affinity
Chromatography, Gel
Humans
MCF-7 Cells
Macrolides
Multiprotein Complexes
Protein Interaction Mapping
Protein Multimerization
Tandem Mass Spectrometry
English Mass spectrometry (MS)-based identification and characterization of protein complexes is becoming a prerequisite for in-depth biochemical analyses of intracellular processes. Here, we describe two state-of-the-art MS-based approaches to characterize protein-protein interactions and multi-protein complexes involved in autophagy in mammalian cells. The combination of affinity purification (AP)-MS, which identifies binary protein-protein interactions, with size-exclusion chromatography (SEC)-protein correlation profiling (PCP), which helps monitor protein complex assemblies, is a powerful tool to acquire a full overview of the interlinkage and regulation of novel multi-protein complexes that might play a role in autophagy.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/93883
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