論文 - 詳細
| RRC ID | 43991 |
|---|---|
| 著者 | Kanno T, Tsuchiya A, Shimizu T, Tanaka A, Nishizaki T. |
| タイトル | Novel indomethacin action: selective and direct activation of protein kinase C-ε. |
| ジャーナル | Cell Physiol Biochem |
| Abstract |
BACKGROUND/AIMS:In our earlier study, indomethacin potentiated α7 acetylcholine (ACh) receptor responses by activating protein kinase C (PKC). The present study was conducted to gain further insight into the indomethacin action on PKC. METHODS:PKC activity was assayed in PC-12 cells or under the cell-free conditions. PKC-ε was knocked-down using the siRNA to silence the PKC-ε-targeted gene. A fluorescein-conjugated indomethacin was synthesized to examine the interaction of indomethacin with PKC-ε. RESULTS:In the in situ PKC assay, indomethacin activated PKC in PC-12 cells in a concentration (1-100 µM)-dependent manner, and the activation was suppressed by knocking-down PKC-ε. In the cell-free PKC assay, indomethacin (100 µM) activated PKC-ε in the absence of diacylglycerol, phosphatidylserine, and calcium, but other PKC isozymes such as α, βΙ, βΙΙ, γ, δ, ι, and ζ were not activated. In the indomethacin binding assay using a fluorescent-conjugated indomethacin on blue native-polyacrylamide gel electrophoresis (blue native-PAGE), a fluorescent signal was detected at the site consistent with PKC-ε protein and the signal was attenuated by adding non-conjugated indomethacin or eliminated by pretreatment with non-conjugated indomethacin. CONCLUSION:The results of the present study show that indomethacin has the potential to selectively activate PKC-ε through its direct binding, independently of cyclooxygenase (COX) inhibition. |
| 巻・号 | 30(3) |
| ページ | 771-7 |
| 公開日 | 2012-1-1 |
| DOI | 10.1159/000341456 |
| PII | 000341456 |
| PMID | 22854271 |
| MeSH | Animals Cyclooxygenase Inhibitors / pharmacology* Enzyme Activation / drug effects* Fluorescein / chemistry Indomethacin / pharmacology* PC12 Cells Prostaglandin-Endoperoxide Synthases / chemistry Prostaglandin-Endoperoxide Synthases / metabolism Protein Binding Protein Isoforms / metabolism Protein Kinase C-epsilon / chemistry Protein Kinase C-epsilon / genetics Protein Kinase C-epsilon / metabolism* RNA Interference RNA, Small Interfering / metabolism Rats Substrate Specificity |
| IF | 5.5 |
| 引用数 | 3 |
| WOS 分野 | PHYSIOLOGY CELL BIOLOGY |
| リソース情報 | |
| ヒト・動物細胞 | PC-12(RCB0009) |