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© The Rockefeller University Press,
0021-9525/2001//165 $5.00
The Journal of Cell Biology, Volume 152, Number 1,
, 2001 165-180
Original Article |
The Phagosome Proteome
: Insight into Phagosome Functions
b Département de Pathologie et Biologie Cellulaire, Université de Montréal, Montréal, Quebec, Canada, H3C 3J7
c Neurodégénérescence et Plasticité, Hopital A. Michallon, Centre Hospitalier Universitaire, 38043 Grenoble, France
Département de pathologie et biologie cellulaire, Université de Montréal, C.P. 6128, Succ. Centre ville, Montréal, Québec, Canada, H3C 3J7.(514) 343-2459(514) 343-7250
michel.desjardins{at}umontreal.ca
Phagosomes are key organelles for the innate ability of macrophages to participate in tissue remodeling, clear apoptotic cells, and restrict the spread of intracellular pathogens. To understand the functions of phagosomes, we initiated the systematic identification of their proteins. Using a proteomic approach, we identified >140 proteins associated with latex bead–containing phagosomes. Among these were hydrolases, proton pump ATPase subunits, and proteins of the fusion machinery, validating our approach. A series of unexpected proteins not previously described along the endocytic/phagocytic pathways were also identified, including the apoptotic proteins galectin3, Alix, and TRAIL, the anti-apoptotic protein 14-3-3, the lipid raft-enriched flotillin-1, the anti-microbial molecule lactadherin, and the small GTPase rab14. In addition, 24 spots from which the peptide masses could not be matched to entries in any database potentially represent new phagosomal proteins. The elaboration of a two-dimensional gel database of >160 identified spots allowed us to analyze how phagosome composition is modulated during phagolysosome biogenesis. Remarkably, during this process, hydrolases are not delivered in bulk to phagosomes, but are instead acquired sequentially. The systematic characterization of phagosome proteins provided new insights into phagosome functions and the protein or groups of proteins involved in and regulating these functions.
Key Words: phagosome • proteome • matrix-assisted laser desorption/ionization–time-of-flight–mass spectrometry • membrane fusion • apoptosis
© 2001 The Rockefeller University Press
Drs. Garin and Diez contributed equally to this work and should be considered co-first authors.Abbreviations used in this paper: 2-D, two-dimensional; ARF, ADP-ribosylation factor; EST, expression sequence tag; MHC, major histocompatibility complex; MS, mass spectrometry; PDI, protein disulfide isomerase; PMF, peptide mass fingerprinting.
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