Altered protein kinase C in a mast cell variant defective in exocytosis

Journal Article · Sun Mar 01 04:00:00 EST 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.5.1277· OSTI ID:6175036

Mazurek, N; Regazzi, R; Borner, C; Wyss, R; Erne, P; Eppenberger, U; Fabbro, D

The murine mast cell line PB-3c is dependent on interleukin 3 (IL-3) with respect to survival and proliferation. These cells also require IL-3 to display antigen-mediated serotonin release, which is coupled to a transient increase of cytosolic free calcium ((Ca/sup 2 +/)/sub i/). The antigen-mediated exocytosis is inhibited by phorbol 12-tetradecanoate 13-acetate (PTA), an activator of phospholipid/Ca/sup 2 +/-sensitive protein kinase. In contrast, the malignant mast cell variant PB-1 is IL-3 independent with respect to proliferation but is unable to undergo antigen-mediated exocytosis. Yet this cell line exhibits basal levels of (Ca/sup 2 +/)/sub i/, serotonin content, and numbers of IgE receptors comparable to those of PB-3c cells. Subcellular distribution studies revealed that the specific activity of cytosolic protein kinase C or PB-1 cells was only 40% of that found in PB-3c cells. Scatchard plot analysis of (/sup 3/H)-phorbol 12,13-dibutyrate binding to intact PB-1 cells demonstrated the presence of 20% high-affinity and 80% low-affinity phorbol ester receptors, whereas PB-3c cells displayed only the low-affinity phorbol ester binding. Immunological characterization of protein kinase C from both cell lines revealed the presence of a normal 77-kDa protein kinase C holoenzyme in both cell lines. In addition, a 72-kDa protein kinase C-related protein band was found mainly in the membrane fraction of the PB-1 variant. It is suggested that this altered and membrane-bound form of protein kinase C may be involved in the blockage of the antigen-mediated exocytosis of PB-1 cells.

• Research Organization: Univ. Clinic Medical School, Basel, Switzerland
• OSTI ID: 6175036
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:5; ISSN PNASA
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKALINE EARTH METAL COMPOUNDS
AMINES
ANIMAL CELLS
ANIMALS
AROMATICS
AUTONOMIC NERVOUS SYSTEM AGENTS
AZAARENES
AZOLES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
CALCIUM COMPOUNDS
CARCINOGENS
CATIONS
CHARGED PARTICLES
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DISTRIBUTION
DRUGS
ELECTROPHORESIS
ENZYMES
ESTERS
GROWTH FACTORS
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
INDOLES
IONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
LYMPHOKINES
MAMMALS
MAST CELLS
MEMBRANE PROTEINS
MICE
MITOGENS
NEUROREGULATORS
NUCLEI
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHORBOL ESTERS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PROTEINS
PYRROLES
RADIOISOTOPES
RADIOPROTECTIVE SUBSTANCES
REACTION KINETICS
RECEPTORS
RODENTS
SEROTONIN
SOMATIC CELLS
SUBCELLULAR DISTRIBUTION
SYMPATHOMIMETICS
TRANSFERASES
TRITIUM COMPOUNDS
TRYPTAMINES
VERTEBRATES