Localization of phospholipase A2 in normal and ras-transformed cells

Bar-Sagi, D., Suhan, J. P., McCormick, F., Feramisco, J. R. (May 1988) Localization of phospholipase A2 in normal and ras-transformed cells. J Cell Biol, 106 (5). pp. 1649-58. ISSN 0021-9525

URL: http://www.ncbi.nlm.nih.gov/pubmed/3286660

Abstract

The cellular localization of phospholipase A2 (PLA2) was examined in normal and ras-transformed rat fibroblasts using immunohistochemical techniques. Polyclonal antibodies were generated against porcine pancreatic PLA2 and were affinity purified for use in this study. The antibodies detected a 16-kD band on immunoblots of total cellular proteins from fibroblasts. In cell-free assays of phospholipase A2 activity, the purified antibodies inhibited the bulk of the enzyme activity whereas control IgG preparations had no effect. Immunofluorescence microscopy indicated that PLA2 was diffusely distributed throughout the cell. Increased concentration of PLA2 was detected under membrane ruffles in normal and ras-transformed cells. Specific immunofluorescence staining was also detected on the outer surface of the normal cells. Immunoelectron microscopy demonstrated the increased accumulation of PLA2 in membrane ruffles and also revealed the presence of the enzyme in microvilli and its association with intracellular vesicles. Ultrastructural localization of PLA2 and the ras oncogene protein, using a double immunogold labeling technique, indicated a spatial proximity between PLA2 and ras proteins in the ruffles of ras-transformed cells. The possible role of PLA2 in the structural rearrangements that underlie membrane ruffling is discussed.

Item Type: Paper
Uncontrolled Keywords: Animals Antibodies/immunology/isolation & purification Cell Line Cell Line, Transformed Cell Membrane/analysis/*enzymology Chromatography, Affinity Cytoplasm/*enzymology Fibroblasts Fluorescent Antibody Technique Immunohistochemistry Intracellular Membranes/analysis/*enzymology Membrane Proteins/analysis Microscopy, Electron Microscopy, Electron, Scanning Microvilli/analysis/enzymology Phospholipases/*analysis Phospholipases A/*analysis/immunology Proto-Oncogene Protein p21(ras) Proto-Oncogene Proteins/*analysis Rats Research Support, U.S. Gov't, P.H.S.
Subjects: organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > phospholipase A
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > G protein > Ras
CSHL Authors:
Communities: CSHL labs
Depositing User: Gail Sherman
Date: 1 May 1988
Date Deposited: 10 Nov 2017 16:49
Last Modified: 10 Nov 2017 16:49
PMCID: PMC2115042
Related URLs:
URI: http://repository.cshl.edu/id/eprint/35118

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