Phorbol myristate acetate receptors in human polymorphonuclear neutrophils
Resting or phorbol myristate acetate (PMA)-pretreated neutrophils were disrupted by nitrogen cavitation and were fractionated on Percoll density gradients to identify the subcellular location of PMA receptors. Receptors were found in the cytoplasm of resting cells; neither primary nor secondary granules bound (/sup 3/H)PMA, and the few binding sites located in non-granule membrane fractions appeared to reflect cytosolic contamination. Contrastingly, PMA-pretreated cells lost cytosolic receptors; > 80% of PMA-binding sites were associated with non-granule membranes. Protein kinase C activity similarly shifted from cytosol to membranes after PMA treatment. Indeed, protein kinase C and PMA receptors co-sedimented on Percoll gradients, co-eluted from Ultragel AcA 44 columns loaded with neutrophil cytoplasm, and were identically influenced by various phospholipids. Finally, PMA, mezerein, diacylglycerol, and dialkylglycerol activated protein kinase C with potencies that paralleled their respective abilities to stimulate neutrophil aggregation responses and inhibit (/sup 3/H)PMA binding to whole cells or cytosol. These results fit a model of stimulus-response coupling wherein exogenous PMA or endogenous diacylglycerol solvate in cellular membranes. Cytosolic protein kinase C binds to the intramembranous ligand, forming an active, membrane-associated complex that phosphorylates nearby elements involved in triggering aggregation and other responses.
- Research Organization:
- Wake Forest Univ. Medical Center, Winston-Salem, NC
- OSTI ID:
- 6935654
- Journal Information:
- J. Immunol.; (United States), Journal Name: J. Immunol.; (United States) Vol. 135:5; ISSN JOIMA
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARCINOGENS
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
ENZYME ACTIVITY
ENZYMES
ESTERS
EXTRACTION CHROMATOGRAPHY
HYDROGEN ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPIDS
MEMBRANE PROTEINS
MEMBRANES
METABOLISM
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
PHORBOL ESTERS
PHOSPHOLIPIDS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTEINS
RADIOISOTOPES
RADIORECEPTOR ASSAY
RECEPTORS
SEDIMENTATION
SEPARATION PROCESSES
TRACER TECHNIQUES
TRANSFERASES
TRITIUM
YEARS LIVING RADIOISOTOPES