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Molecular & Cellular Proteomics 5:2228-2243, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Proteomics Fingerprinting of Phagosome Maturation and Evidence for the Role of a G during Uptake^*,S

From the ^a Department of Molecular Cell Research, Max Planck Institute for Medical Research, ^b Department of Internal Medicine IV, University Hospital of Heidelberg, and ^e Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), D-69120 Heidelberg, Germany, ^d Department of Biological Sciences, Imperial College, London SW7 2AZ, United Kingdom, ^f ISREC National Centre of Competence in Research (NCCR) Molecular Oncology, Swiss Institute of Experimental Cancer Research (ISREC), Epalinges, CH-1006 Switzerland, ^g ISREC Swiss Institute of Bioinformatics (SIB), CH-1015 Lausanne, Switzerland, and ⁱ Départment de Biochimie, Faculté des Sciences, Université de Genève, CH-1211 Genève-4, Switzerland

Phagocytosis, whether of food particles in protozoa or bacteria and cell remnants in the metazoan immune system, is a conserved process. The particles are taken up into phagosomes, which then undergo complex remodeling of their components, called maturation. By using two-dimensional gel electrophoresis and mass spectrometry combined with genomic data, we identified 179 phagosomal proteins in the amoeba Dictyostelium, including components of signal transduction, membrane traffic, and the cytoskeleton. By carrying out this proteomics analysis over the course of maturation, we obtained time profiles for 1,388 spots and thus generated a dynamic record of phagosomal protein composition. Clustering of the time profiles revealed five clusters and 24 functional groups that were mapped onto a flow chart of maturation. Two heterotrimeric G protein subunits, G₄ and Gß, appeared at the earliest times. We showed that mutations in the genes encoding these two proteins produce a phagocytic uptake defect in Dictyostelium. This analysis of phagosome protein dynamics provides a reference point for future genetic and functional investigations.

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