Lyn and Syk kinases are sequentially engaged in phagocytosis mediated by Fc gamma R.
Recent data indicate that phagocytosis mediated by FcgammaRs is controlled by the Src and Syk families of protein tyrosine kinases. In this study, we demonstrate a sequential involvement of Lyn and Syk in the phagocytosis of IgG-coated particles. The particles isolated at the stage of their binding to FcgammaRs (4 degrees C) were accompanied by high amounts of Lyn, in addition to the signaling gamma-chain of FcgammaRs. Simultaneously, the particle binding induced rapid tyrosine phosphorylation of numerous proteins. During synchronized internalization of the particles induced by shifting the cell to 37 degrees C, Syk kinase and Src homology 2-containing tyrosine phosphatase-1 (SHP-1) were associated with the formed phagosomes. At this step, most of the proteins were dephosphorylated, although some underwent further tyrosine phosphorylation. Quantitative immunoelectron microscopy studies confirmed that Lyn accumulated under the plasma membrane beneath the bound particles. High amounts of the gamma-chain and tyrosine-phosphorylated proteins were also observed under the bound particles. When the particles were internalized, the gamma-chain was still detected in the region of the phagosomes, while amounts of Lyn were markedly reduced. In contrast, the vicinity of the phagosomes was heavily decorated with anti-Syk and anti-SHP-1 Abs. The local level of protein tyrosine phosphorylation was reduced. The data indicate that the accumulation of Lyn during the binding of IgG-coated particles to FcgammaRs correlated with strong tyrosine phosphorylation of numerous proteins, suggesting an initiating role for Lyn in protein phosphorylation at the onset of the phagocytosis. Syk kinase and SHP-1 phosphatase are mainly engaged at the stage of particle internalization.
Strzelecka-Kiliszek
Agnieszka
A
Department of Cell Biology, Nencki Institute of Experimental Biology, Warsaw, Poland.
Kwiatkowska
Katarzyna
K
Sobota
Andrzej
A
D000818:Animals; D002460:Cell Line; D004792:Enzyme Precursors / Q000032:analysis / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D047908:Intracellular Signaling Peptides and Proteins; D008264:Macrophages / Q000737:chemistry / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D016253:Microscopy, Immunoelectron; D010587:Phagocytosis / Q000276:immunology*; D019000:Phosphotyrosine / Q000032:analysis / Q000378:metabolism; D054645:Protein Phosphatase 1; D021381:Protein Transport / Q000276:immunology; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D011505:Protein-Tyrosine Kinases / Q000032:analysis / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D017452:Receptors, IgG / Q000032:analysis / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D015398:Signal Transduction / Q000276:immunology; D000072377:Syk Kinase; D019061:src-Family Kinases / Q000032:analysis / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure
D004792:Enzyme Precursors; D047908:Intracellular Signaling Peptides and Proteins; D017452:Receptors, IgG; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D000072377:Syk Kinase; C000606897:Syk protein, mouse; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D054645:Protein Phosphatase 1; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases; C509248:Ptpn6 protein, mouse