Influence of calcium on the subcellular distribution of protein kinase C in human neutrophils. Extraction conditions determine partitioning of histone-phosphorylating activity and immunoreactivity between cytosol and particulate fractions.
Activation of the neutrophil respiratory burst is thought to involve a translocation and activation of protein kinase C. We report that the presence of Ca2+ during the disruption of unstimulated human neutrophils and cytoplasts resulted in an increase in protein kinase C activity (histone phosphorylation) and immunoreactive protein kinase C species in the particulate (membrane) fraction and a reduction in such activities in the cytosol. This Ca2+-induced translocation of activity was concentration-dependent and occurred at physiologically relevant concentrations of Ca2+ (30-500 nM). The Ca2+-induced membrane association of protein kinase C could be reversed by removal of Ca2+. These findings indicate that the Ca2+ concentration of the extraction buffer can determine the subcellular distribution of protein kinase C in disrupted cells and suggest that the observed location of this enzyme activity in cell fractions may not necessarily reflect the localization in intact cells. These results also raise the possibility that the distribution of protein kinase C between cytosol and membrane is a dynamic equilibrium controlled by levels of free Ca2+. Thus, Ca2+ might regulate distribution as well as activation of protein kinase C.
Phillips
W A
WA
Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia 19104.
Fujiki
T
T
Rossi
M W
MW
Korchak
H M
HM
Johnston
R B
RB
D001692:Biological Transport; D002118:Calcium / Q000494:pharmacology*; D002462:Cell Membrane / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology*; D004075:Diglycerides / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D008970:Molecular Weight; D009504:Neutrophils / Q000187:drug effects / Q000648:ultrastructure*; D010718:Phosphatidylserines / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D004075:Diglycerides; D010718:Phosphatidylserines; D004533:Egtazic Acid; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium