1. Proteomic analysis reveals a role for protein kinase C-α in phagosome maturation
- Author
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Albert Descoteaux, John Derek Ng Yan Hing, Michel Desjardins, Institut Armand Frappier (INRS-IAF), Institut National de la Recherche Scientifique [Québec] (INRS)-Réseau International des Instituts Pasteur (RIIP), and Université de Montréal (UdeM)
- Subjects
Proteomics ,Proteome ,Macrophage ,MESH: Cathepsin D ,Endocytic cycle ,Cathepsin D ,Biochemistry ,Mice ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,Phagosomes ,MESH: Animals ,MESH: Antigens, CD ,Protein Kinase C ,Phagosome ,Microspheres ,Cell biology ,MESH: Proteome ,MESH: Protein Kinase C-alpha ,MESH: Membrane Proteins ,MESH: Mutation ,Protein Kinase C-alpha ,MESH: Lysosome-Associated Membrane Glycoproteins ,Endosome ,Phagocytosis ,MESH: Microspheres ,Biophysics ,Biology ,Phagolysosome ,Cell Line ,MESH: Phagosomes ,Antigens, CD ,Phagosome maturation ,Animals ,MESH: Mice ,Molecular Biology ,Protein kinase C ,Macrophages ,MESH: Biological Markers ,MESH: Macrophages ,Lysosome-Associated Membrane Glycoproteins ,Membrane Proteins ,rab7 GTP-Binding Proteins ,Cell Biology ,MESH: Protein Kinase C ,Cathepsins ,MESH: Cell Line ,MESH: rab GTP-Binding Proteins ,rab GTP-Binding Proteins ,MESH: Cathepsins ,Mutation ,Lysosomes ,Biomarkers ,MESH: Lysosomes - Abstract
International audience; Acquisition of microbicidal properties by phagosomes requires the action of molecules which regulate the interactions between phagosomes and endocytic organelles. Members of the protein kinase C (PKC) superfamily of serine/threonine kinases are recruited to phagosomes with various kinetics during phagolysosome biogenesis. To study the role of PKC-alpha in this process, we compared the composition of latex bead-containing phagosomes isolated from control and dominant-negative (DN) PKC-alpha-overexpressing RAW 264.7 macrophages. Western blot analysis indicated that the levels of both lysosomal-associated membrane protein-1 and flotillin-1, which are acquired through interactions with late endosomes and lysosomes, are reduced in phagosomes from DN PKC-alpha-overexpressing macrophages. Proteomic characterization of latex bead-containing phagosomes revealed that recruitment of the small GTPase Rab7, cathepsin D, and cathepsin S is inhibited by DN PKC-alpha. Collectively, these data provide evidence that PKC-alpha plays a role in phagolysosome biogenesis, a critical process of the innate immune response against infections.
- Published
- 2004
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