acid-phosphatase and Mast-Cell-Sarcoma

The 3H-labeled prostaglandin D2 [( 3H]PGD2) binding protein in the membrane fraction of mastocytoma P-815 cells was characterized. The specific binding of [3H]PGD2 to the cells or the membranes reached a maximum at pH 5.6, and was saturable, displaceable and of high affinity when incubated at 0 or 37 degrees C. The Bmax values for [3H]PGD2 binding in the two preparations at pH 5.6 were much higher at 0 degrees C than at 37 degrees C, whereas the Kd values were almost equal (85.3 nM for the cells and 80.5 nM for the membranes, respectively). High specific [3H]PGD2 binding activity in the mildly acid-treated cells was still observed when the external pH was raised from 5.6 to 7.2. Furthermore, specific [3H]PGD2 binding to the membranes (at 0 degrees C, pH 5.6) increased on addition of phosphatase inhibitors (NaF and molybdate) in the presence of 10 microM ATP, but practically disappeared on pretreatment of the membranes with phosphatase. On incubation of the membrane with [gamma-32P]ATP and molybdate, the stimulated incorporation of the [32P]phosphate into several peptides, including ones having an Mr of around 100,000-120,000, was observed. These results suggest that [3H]PGD2 binding in the mastocytoma P-815 cell membrane is controlled through phosphorylation-dephosphorylation of the receptor itself.

Topics: Acid Phosphatase; Animals; Cell Line; Cell Membrane; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Kinetics; Male; Mast-Cell Sarcoma; Mice; Molybdenum; Phosphorylation; Prostaglandin D2; Receptors, Immunologic; Receptors, Prostaglandin; Sodium Fluoride; Subcellular Fractions; Temperature

The phospholipids from murine mastocytoma FMA3 and P-815 clone cells were quantitatively analyzed, and the major glycerophospholipids were examined for their fatty acyl chain distribution. In these cells, the content of histamine was less than 1/100 of normal mouse mast cells, and FMA3 cells had 1.5-fold as much histamine content as P-815 cells. The predominant phospholipid species of both mastocytoma FMA3 and P-815 were choline-containing glycerophospholipids (48%) and ethanolamine-containing glycerophospholipids (29%). The remaining minor constituents were sphingomyelin (6%, 7%), phosphatidylinositol (7%, 5%), phosphatidylserine (2%, 5%), cardiolipin (4%, 3%), and phosphatidic acid (2%, 1% for FMA3 and P-815, respectively). The choline-containing glycerophospholipids consisted of high amounts of 1-O-alkyl-2-acyl type (31%, 25%) and 1,2-diacyl type (63%, 66%) and a smaller amount of 1-O-alk-1'-enyl-2-acyl type (7%, 8%). In contrast, ethanolamine-containing glycerophospholipids were characterized by high contents of 1-O-alk-1'-enyl-2-acyl type (36%, 31%) and 1,2-diacyl type (55%, 58%), and a lower level of 1-O-alkyl-2-acyl type (12% and 11% for FMA3 and P-815, respectively). Unlike choline-containing glycerophospholipids and sphingomyelin that were rich in palmitic acid, ethanolamine-containing glycerophospholipids, phosphatidylserine and phosphatidylinositol showed a high proportion of stearic acid in the overall fatty acid composition. The content of arachidonic acid was highest in phosphatidylinositol. Sphingomyelin had a large amount of long chain and polyunsaturated fatty acids. In both choline- and ethanolamine-containing glycerophospholipids, the predominant fatty acids in the sn-1-position were palmitic, stearic, and oleic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Cell Line; Clone Cells; Fatty Acids; Histamine; Mast-Cell Sarcoma; Mice; Phospholipids

H-2b lymphocytes were sensitized against H-2d alloantigens by lymphocyte culture reaction and incubated with H-2d mastocytoma cells. The interaction between lymphoid cells and mastocytoma cells was stopped by fixation with glutaraldehyde. The areal density of the cytoplasmic vacuoles as well as the subcellular localization of acid phosphatase in lymphocytes were examined by electron microscopy. Two populations of lymphocytes were observed, small lymphocytes with heterochromatic nuclei and larger lymphocytes (lymphoblasts) with euchromatic nuclei. Only the lymphoblasts showed change following interaction with target cells. The vacuole area in percent of cytoplasmic area (vacuole areal density) of sensitized lymphoblasts increased during the first 30 minutes and from the third to fourth hour of interaction with target cells. Acid phosphatase staining was observed in the Golgi apparatus of the lymphoblasts after 30 minutes of interaction. Multivesicular bodies showed acid phosphatase staining within 20 minutes of interaction with target cells. After 20 minutes of interaction, phagosomes containing myelin figures were formed. These phagosomes also showed acid phosphatase staining and during the next hours of interaction their number increased over the number of multivesicular bodies.

Topics: Acid Phosphatase; Animals; Cell Count; Cytoplasm; Golgi Apparatus; Guinea Pigs; Isoantigens; Lymphocyte Culture Test, Mixed; Lymphocytes; Lysosomes; Mast-Cell Sarcoma; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Organoids; Vacuoles

Topics: Acid Phosphatase; Adenosine Triphosphatases; Alkaline Phosphatase; Animals; Bone Marrow; Connective Tissue; Dog Diseases; Dogs; Enzymes; Glucosephosphate Dehydrogenase; Glutamate Dehydrogenase; Histocytochemistry; L-Lactate Dehydrogenase; Leukemia; Malate Dehydrogenase; Mast-Cell Sarcoma; Succinate Dehydrogenase

Topics: Acid Phosphatase; Animals; Autoradiography; Dogs; Dopa Decarboxylase; Humans; Mast Cells; Mast-Cell Sarcoma; Melanins; Mice; Microscopy, Electron; Nevus, Pigmented; Oxidoreductases; Peritoneum; Peroxidases; Rats; Skin; Skin Neoplasms; Urticaria Pigmentosa