Autophagosomal proteome analysis by protein correlation profiling-SILAC

Methods Mol Biol. 2014:1188:271-9. doi: 10.1007/978-1-4939-1142-4_19.

Abstract

Autophagy is one of the two major degradation pathways within eukaryotic cells. Nevertheless, little is known about the protein composition of autophagosomes, the vesicles shuttling proteins to lysosomes for degradation. Protein correlation profiling in combination with stable isotope labeling by amino acids in cell culture is a stringent method to investigate the dynamics of the autophagosomal proteome. It enables the discrimination between autophagosomal and co-purifying proteins identifying organellar candidate proteins for further investigation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry*
  • Analytic Sample Preparation Methods
  • Cell Fractionation
  • Cells, Cultured
  • Chromatography, Liquid
  • Isotope Labeling / methods*
  • Mass Spectrometry
  • Phagosomes / metabolism*
  • Protein Transport
  • Proteins / chemistry*
  • Proteins / metabolism*
  • Proteomics / methods*

Substances

  • Amino Acids
  • Proteins