Activation of the respiratory burst in macrophages. Phosphorylation specifically associated with Fc receptor-mediated stimulation.
Inflammatory macrophages elicited from the peritoneal cavity of mice injected with endotoxin can avidly ingest E opsonized with IgG antibody (EIgG) or with IgM antibody and C (EIgMC). However, only ingestion of EIgG is associated with activation of the respiratory burst and release of superoxide anion. We compared the endogenous phosphorylation of proteins from macrophages stimulated by interaction with EIgG or EIgMC on the premise that proteins phosphorylated after stimulation by EIgG but not EIgMC could play a role in activating the enzyme (oxidase) responsible for the respiratory burst. Proteins were separated by one-dimensional and two-dimensional electrophoresis in polyacrylamide gels. We found that proteins with approximate Mr of 20 kDa, 23 kDa, 46 kDa, 48 kDa (three proteins), 67 kDa, and 130 kDa were more heavily phosphorylated after EIgG stimulation than after EIgMC stimulation. Exposure to PMA, which activates the respiratory burst oxidase, induced phosphorylation of the 23-kDa, 48-kDa group, and 130-kDa proteins that were phosphorylated after stimulation by EIgG. Activity of protein kinase C was found to be significantly increased in the particulate fraction of macrophages stimulated by EIgG but not in the particulate fraction of EIgMC-stimulated cells. These data are compatible with the hypotheses that phosphorylation of specific cellular proteins, especially with a Mr of approximately 48 kDa, is involved in activation of the respiratory burst oxidase, and that function of protein kinase C also plays a part in this activation process.
Brozna
J P
JP
Department of Pathology, Denver Veterans Administration Medical Center, CO 80220.
Hauff
N F
NF
Phillips
W A
WA
Johnston
R B
RB
D000818:Animals; D003165:Complement System Proteins / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D008262:Macrophage Activation; D008264:Macrophages / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010101:Oxygen Consumption; D010587:Phagocytosis; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011961:Receptors, Fc / Q000502:physiology*; D013347:Subcellular Fractions / Q000201:enzymology
D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011961:Receptors, Fc; D003165:Complement System Proteins; D011493:Protein Kinase C