Assessing the phagosome proteome by quantitative mass spectrometry

Julien Peltier, Anetta Härtlova, Matthias Trost (Lead / Corresponding author)

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

3 Citations (Scopus)

Abstract

Phagocytosis is the process that engulfs particles in vesicles called phagosomes that are trafficked through a series of maturation steps, culminating in the destruction of the internalized cargo. Because phagosomes are in direct contact with the particle and undergo constant fusion and fission events with other organelles, characterization of the phagosomal proteome is a powerful tool to understand mechanisms controlling innate immunity as well as vesicle trafficking. The ability to isolate highly pure phagosomes through the use of latex beads led to an extensive use of proteomics to study phagosomes under different stimuli. Thousands of different proteins have been identified and quantified, revealing new properties and shedding new light on the dynamics and composition of maturing phagosomes and innate immunity mechanisms. In this chapter, we describe how quantitative-based proteomic methods such as label-free, dimethyl labeling or Tandem Mass Tag (TMT) labeling can be applied for the characterization of protein composition and translocation during maturation of phagosomes in macrophages.

Original languageEnglish
Title of host publicationPhagocytosis and phagosomes
Subtitle of host publicationmethods and protocols
EditorsRoberto Botelho
Place of PublicationNew York
PublisherSpringer
Pages249-263
Number of pages15
ISBN (Electronic)9781493965816
ISBN (Print)9781493965793
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in molecular biology
PublisherSpringer
Volume1519
ISSN (Print)1064-3745

Keywords

  • Maturation of phagosomes
  • Macrophages
  • Label free
  • Isotope labeling
  • Quantitative proteomics
  • Phagosome dynamics
  • Dimethyl
  • TMT
  • Mass spectrometry

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