
Ligation of the alpha2M* signaling receptor regulates synthesis of cytosolic phospholipase A2.
We have studied the regulation of cytosolic phospholipase A2 (cPLA2) synthesis in macrophages stimulated with receptor-recognized forms of alpha2-macroglobulin (alpha2M*). [35S]methionine-labeled cells were stimulated with alpha2M* and [35S]cPLA2 was immunoprecipitated with a monoclonal antibody directed against cPLA2. The precipitates were electrophoresed, immunoblotted, cPLA2 detected by Enhanced Chemifluorescence, and its radioactivity determined. Stimulation of cells with alpha2M* caused a two- to threefold increase in cPLA2 synthesis compared to buffer-treated cells which was consistently maximal at 200 pM of alpha2M*. Actinomycin D or cycloheximide treatment of cells drastically reduced alpha2M*-induced cPLA2 synthesis. Likewise, inhibition of protein kinase C with chelerythrin, farnesyl transferase with manumycin A, MEK kinase with U0126, Erk1/2 kinases with PD98059, p38MAPK with SB203580, PI 3-kinase with wortmannin or LY294002, p70s6k with rapamycin, or depletion of [Ca2+]i with either BAPTA/AM or EGTA drastically reduced alpha2M* induction of cPLA2. Inhibition of NFKB activation with BAY11-7182 or PGA1 also abolished alpha2M* induction of cPLA2. We conclude that alpha2M*-induced cPLA2 synthesis is controlled by [Ca2+]i levels, tyrosine kinase activity, the p21ras-dependent MAPK and PI 3-kinase downstream signaling pathways, and regulation of NFkappaB.
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Related Subject Headings
- alpha-Macroglobulins
- Signal Transduction
- Ribosomal Protein S6 Kinases
- Receptors, Immunologic
- Proto-Oncogene Proteins p21(ras)
- Protein-Tyrosine Kinases
- Protein Serine-Threonine Kinases
- Protein Kinase C
- Phospholipases A2
- Phospholipases A
Citation

Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- alpha-Macroglobulins
- Signal Transduction
- Ribosomal Protein S6 Kinases
- Receptors, Immunologic
- Proto-Oncogene Proteins p21(ras)
- Protein-Tyrosine Kinases
- Protein Serine-Threonine Kinases
- Protein Kinase C
- Phospholipases A2
- Phospholipases A