Translocation of protein kinase C to subcellular fractions of human neutrophils.
The subcellular localization of protein kinase C in unstimulated human neutrophils and neutrophils stimulated by phorbol-myristate-acetate (PMA), 1-oleoyl-2-acetyl-rac-glycerol (OAG), and ionomycin was investigated in subcellular fractions obtained by nitrogen cavitation and Percoll density gradient centrifugation. Protein kinase C was found to be localized mainly in the cytosol in unstimulated cells, whereas significant translocation to fractions containing the plasma membrane was observed after stimulation by PMA, OAG, and ionomycin. At the same time, phospholipid-insensitive protein kinase activity appeared in the cytosol and the plasma membrane fractions. To determine whether binding of protein kinase C occurred to the plasma membrane or to intracellular membranes that had translocated to the plasma membrane, we investigated the ability of isolated azurophil, specific and secretory granules, and plasma membrane vesicles to bind protein kinase C in response to addition of PMA and OAG. Only fractions containing plasma membranes and secretory granules were able to bind protein kinase C. The observation explains the selective activation of plasma membrane structures by protein kinase C.
Christiansen
N O
NO
Department of Medicine and Infectious Diseases, Marselisborg Hospital, Aarhus, Denmark.
Borregaard
N
N
D001665:Binding Sites / Q000187:drug effects; D003594:Cytoplasmic Granules / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004987:Ethers / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects; D006801:Humans; D015759:Ionomycin; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology*; D011493:Protein Kinase C / Q000378:metabolism* / Q000502:physiology; D013347:Subcellular Fractions / Q000201:enzymology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; D004987:Ethers; D015759:Ionomycin; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate