Unraveling the human dendritic cell phagosome proteome by organellar enrichment ranking

Buschow, Sonja I., Lasonder, Edwin, Szklarczyk, Radek, Oud, Machteld M., de Vries, I. Jolanda M. and Figdor, Carl G. (2012) Unraveling the human dendritic cell phagosome proteome by organellar enrichment ranking. Journal of Proteomics, 75 (5). pp. 1547-1562. ISSN 1874-3919

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Official URL: https://doi.org/10.1016/j.jprot.2011.11.024


Dendritic cells (DC) take up pathogens through phagocytosis and process them into protein and lipid fragments for presentation to T cells. So far, the proteome of the human DC phagosome, a detrimental compartment for antigen processing and presentation as well as for DC activation, remains largely uncharacterized. Here we have analyzed the protein composition of phagosomes from human monocyte-derived DC. For LC-MS/MS analysis we purified phagosomes from DC using latex beads targeted to DC-SIGN, and quantified proteins using a label-free method. We used organellar enrichment ranking (OER) to select proteins with a high potential to be relevant for phagosome function. The method compares phagosome protein abundance with protein abundance in whole DC. Phagosome enrichment indicates specific recruitment to the phagosome rather than co-purification or passive incorporation. Using OER we extracted the most enriched proteins that we further complemented with functionally associated proteins to define a set of 90 phagosomal proteins that included many proteins with established relevance on DC phagosomes as well as high potential novel candidates. We already experimentally confirmed phagosomal recruitment of Galectin-9, which has not been previously associated with phagocytosis, to both bead and pathogen containing phagosomes, suggesting a role for Galectin-9 in DC phagocytosis.

Item Type: Article
Additional Information: Funding Information: The authors would like to thank the Nijmegen Proteomics Facility for usage of the MS instrumentation. R.S. was supported by Horizon grant ( 050-71-053 ) from the Netherlands Organization for Scientific Research (NWO). This work was supported by the Netherlands Organization for Scientific Research (NWO) grant, number 9120.6030 .
Uncontrolled Keywords: Antigen presentation, Dendritic cell, Galectin-9, Phagocytosis, Phagosome interaction network, Proteomics
Subjects: C100 Biology
C500 Microbiology
C700 Molecular Biology, Biophysics and Biochemistry
Department: Faculties > Health and Life Sciences > Applied Sciences
Depositing User: Rachel Branson
Date Deposited: 08 Jun 2021 12:50
Last Modified: 31 Jul 2021 11:04
URI: http://nrl.northumbria.ac.uk/id/eprint/46382

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