PKCalpha regulates phosphorylation and enzymatic activity of cPLA2 in vitro and in activated human monocytes
| Autor: | Rozina Aamir, Venkita Subbulakshmi, Martha K. Cathcart, Claudine M. Oldfield, Crystal M. Weyman, Alan Wolfman, Qing Li |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Protein Kinase C-alpha
Phospholipase Biology Gene Expression Regulation Enzymologic Monocytes Phospholipases A Oligodeoxyribonucleotides Antisense chemistry.chemical_compound Enzyme activator medicine Humans Protein phosphorylation Phosphorylation Protein kinase C Cells Cultured Arachidonic Acid Superoxide Monocyte Cell Biology respiratory system Molecular biology Enzyme Activation medicine.anatomical_structure chemistry lipids (amino acids peptides and proteins) Arachidonic acid |
| Zdroj: | Cellular signalling. 19(2) |
| ISSN: | 0898-6568 |
| Popis: | Phospholipases A(2) (PLA(2)) are potent regulators of the inflammatory response. We have observed that Group IV cPLA(2) activity is required for the production of superoxide anion (O(2)(-)) in human monocytes [Li Q., Cathcart M.K. J. Biol. Chem. 272 (4) (1997) 2404-2411.]. We have previously identified PKCalpha as a kinase pathway required for monocyte O(2)(-) production [Li Q., Cathcart M.K. J. Biol. Chem. 269 (26) (1994) 17508-17515.]. We therefore investigated the potential interaction between PKCalpha and cPLA(2) by evaluating the requirement for specific PKC isoenzymes in the process of activating cPLA(2) enzymatic activity and protein phosphorylation upon monocyte activation. We first showed that general PKC inhibitors and antisense oligodeoxyribonucleotides (ODN) to the cPKC group of PKC enzymes inhibited cPLA(2) activity. To distinguish between PKCalpha and PKCbeta isoenzymes in regulating cPLA(2) protein phosphorylation and enzymatic activity, we employed our previously characterized PKCalpha or PKCbeta isoenzyme-specific antisense ODN [Li Q., Subbulakshmi V., Fields A.P., Murray, N.R., Cathcart M.K., J. Biol. Chem. 274 (6) (1999) 3764-3771]. Suppression of PKCalpha expression, but not PKCbeta expression, inhibited cPLA(2) protein phosphorylation and enzymatic activity. Additional studies ruled out a contribution by Erk1/2 to cPLA(2) phosphorylation and activation. We also found that cPLA(2) co-immunoprecipitated with PKCalpha and vice versa. In vitro studies demonstrated that PKCalpha could directly phosphorylate cPLA(2).and enhance enzymatic activity. Finally, we showed that addition of arachidonic acid restored the production of O(2)(-) in monocytes defective in either PKCalpha or cPLA(2) expression. Taken together, our data suggest that PKCalpha, but not PKCbeta, is the predominant cPKC isoenzyme required for cPLA(2) protein phosphorylation and maximal induction of cPLA(2) enzymatic activity upon activation of human monocytes. Our data also support the concept that the requirements for PKCalpha and cPLA(2) in O(2)(-) generation are solely due to their seminal role in generating arachidonic acid. |
| Databáze: | OpenAIRE |
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