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Contribution of Covalent Protein Modification to the Antiinflammatory Effects of Cyclopentenone Prostaglandins
Author(s) -
PÉREZSALA DOLORES,
CERNUDAMOROLLÓN EVA,
PINEDAMOLINA ESTELA,
CAÑADA F. JAVIER
Publication year - 2002
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.2002.tb04695.x
Subject(s) - biotinylation , chemistry , biochemistry , cysteine , dna , prostaglandin , cyclopentenone , covalent bond , glutathione , transcription factor , microbiology and biotechnology , biology , gene , enzyme , stereochemistry , organic chemistry
A bstract : Cyclopentenone prostaglandins, which are produced during inflammatory processes, may exert a negative feedback on inflammation. These reactive compounds may form covalent adducts with thiol groups in glutathione or in proteins. The transcription factor NF‐κB is key for the expression of numerous proinflammatory genes. We have observed that treatment of mesangial cells with 15‐deoxy‐Δ 12,14 ‐prostaglandin J 2 (15d‐PGJ 2 ) inhibits the cytokine‐elicited DNA binding activity of NF‐κB, both in intact cells and in isolated nuclear extracts, thus suggesting a direct effect on DNA binding. By using a biotinylated 15d‐PGJ 2 derivative, we have observed that 15d‐PGJ 2 forms an adduct with the p50 subunit of NF‐κB, as shown by Western blot and detection with horseradish peroxidase‐conjugated streptavidin. In contrast, a p50 construct that bears a mutation in the cysteine residue involved in DNA binding (Cys62Ser) and is not susceptible to inhibition by 15d‐PGJ 2 does not incorporate biotinylated 15d‐PGJ 2 . The labeling of several polypeptides after incubation of cells with biotinylated 15d‐PGJ 2 suggests that there may be multiple targets for modification by 15d‐PGJ 2 . We propose that the covalent modification of NF‐κB (and potentially other proteins) by 15d‐PGJ 2 may contribute to the antiinflammatory effects of this prostaglandin.