Identification and regulation of protein kinase C-delta in human neutrophils.
The intracellular mechanisms that regulate the function of human neutrophils are not well understood. Receptor-initiated signaling events result in the production of several second messengers (e.g., Ca2+, diacylglycerol, phosphatidic acid, and arachidonic acid) with the potential to activate members of the protein kinase C (PKC) family of signaling enzymes. The mixture of second messenger signaling molecules produced usually varies, depending on the particular receptor engaged. Previous work suggests that PKC has complex regulatory effects on neutrophil function. This may be due to the presence of multiple isoforms of the enzyme family, responding differentially to the second messengers produced. In studies to identify the PKC isoforms present in human neutrophils, we discovered the presence of the PKC isoform delta in these cells. Like other previously identified isoforms (alpha, beta I, beta II, and zeta), delta is a cytosolic enzyme in unstimulated neutrophils and partially translocates to membrane-containing fractions in cells stimulated by either the PKC activator PMA or the chemoattractant FMLP. Partial purification of cytosolic PKC gave two peaks of activity. The beta isoforms predominated in peak I, while the delta isoform predominated in peak II. The identification of delta indicates that neutrophils contain at least one member of the Ca(2+)-independent, diacylglycerol-dependent subfamily of PKC isoforms. Thus, this isoform may participate in Ca(2+)-independent, but diacylglycerol-dependent signaling events in these cells.
Department of Biochemistry, Wake Forest University Medical Center, Winston-Salem, NC 27157, USA.
J D
JD
Kent
S
S
Sergeant
D J
DJ
Burns
L C
LC
McPhail
D001692:Biological Transport; D002118:Calcium / Q000502:physiology; D004075:Diglycerides / Q000502:physiology; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology*; D011493:Protein Kinase C / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism*; D051745:Protein Kinase C-delta; D015398:Signal Transduction / Q000502:physiology
D004075:Diglycerides; D007527:Isoenzymes; C497258:PRKCD protein, human; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D002118:Calcium