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EnglishThe exposure of human endothelial cells to 3-morpholinosydnonimine (SIN-1) induced the expression of cyclooxygenase-2 (COX-2) in a dose-and time-dependent manner. Interestingly, after a prolonged incubation (>8 h) several proteoforms were visualized by Western blot, corresponding to different states of glycosylation of the protein. This effect was specific for SIN-1 that generates peroxynitrite and it was not detected with other nitric oxide-donors. Metabolic labeling experiments using35S or cycloheximide suggested that the formation of hypoglycosylated COX-2 was dependent on de novo synthesis of the protein rather than the deglycosylation of the native protein. Moreover, SIN-1 reduced the activity of the hexokinase, the enzyme responsible for the first step of glycolysis. The hypoglycosylated COX-2 induced by SIN-1 showed a reduced capacity to generate prostaglandins and the activity was only partially recovered after immunoprecipitation. Finally, hypoglycosylated COX-2 showed a more rapid rate of degradation compared to COX-2 induced by IL-1α and an alteration in the localization with an accumulation mainly detected in the nuclear membrane. Our results have important implication to understand the effect of peroxynitrite on COX-2 expression and activity, and they may help to identify new pharmacological tools direct to increase COX-2 degradation or to inhibit its activity.
Cyclooxygenase-2 glycosylation is affected by peroxynitrite in endothelial cells: Impact on enzyme activity and degradation / S. Eligini, S. Colli, A. Habib, G. Aldini, A. Altomare, C. Banfi. - In: ANTIOXIDANTS. - ISSN 2076-3921. - 10:3(2021 Mar 23), pp. 496.1-496.18. [10.3390/antiox10030496]
| Titolo: | Cyclooxygenase-2 glycosylation is affected by peroxynitrite in endothelial cells: Impact on enzyme activity and degradation | |
| Autori: | COLLI, SUSANNA (Secondo) ALTOMARE, ALESSANDRA ANNA (Penultimo) BANFI, CRISTINA (Ultimo) | |
| Parole Chiave: | Cyclooxygenase-2; Endothelial cell; N-linked glycosylation; SIN-1; | |
| Settore Scientifico Disciplinare: | Settore CHIM/01 - Chimica Analitica Settore CHIM/08 - Chimica Farmaceutica | |
| Data di pubblicazione: | 23-mar-2021 | |
| Rivista: | ||
| Tipologia: | Article (author) | |
| Digital Object Identifier (DOI): | http://dx.doi.org/10.3390/antiox10030496 | |
| Appare nelle tipologie: | 01 - Articolo su periodico |
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|---|---|---|---|---|
| antioxidants-10-00496-v2.pdf | Publisher's version/PDF | Open Access Visualizza/Apri |
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