TLR-4 and Sustained Calcium Agonists Synergistically Produce Eicosanoids Independent of Protein Synthesis in RAW264.7 Cells
| dc.contributor.author | Buczynski, Matthew W. | en |
| dc.contributor.author | Stephens, Daren L. | en |
| dc.contributor.author | Bowers-Gentry, Rebecca C. | en |
| dc.contributor.author | Grkovich, Andrej | en |
| dc.contributor.author | Deems, Raymond A. | en |
| dc.contributor.author | Dennis, Edward A. | en |
| dc.contributor.department | School of Neuroscience | en |
| dc.date.accessioned | 2017-01-14T04:42:08Z | en |
| dc.date.available | 2017-01-14T04:42:08Z | en |
| dc.date.issued | 2007-08-03 | en |
| dc.description.abstract | Arachidonic acid is released by phospholipaseA2 and converted into hundreds of distinct bioactive mediators by a variety of cyclooxygenases (COX), lipoxygenases (LO), and cytochrome P450s. Because of the size and diversity of the eicosanoid class of signaling molecules produced, a thorough and systematic investigation of these biological processes requires the simultaneous quantitation of a large number of eicosanoids in a single analysis. We have developed a robust liquid chromatography/tandem mass spectrometry method that can identify and quantitate over 60 different eicosanoids in a single analysis, and we applied it to agonist stimulated RAW264.7 murine macrophages. Fifteen different eicosanoids produced through COX and 5-LO were detected either intracellularly or in the media following stimulation with 16 different agonists of Toll-like receptors (TLR), G protein-coupled receptors, and purinergic receptors. No significant differences in the COX metabolite profiles were detected using the different agonists; however, we determined that only agonists creating a sustained Ca2<sup>2+</sup> influx were capable of activating the 5-LO pathway in these cells. Synergy between Ca2<sup>2+</sup> and TLR pathways was detected and discovered to be independent of NF-κB-induced protein synthesis. This demonstrates that TLR induction of protein synthesis and priming for enhanced phospholipase A<sub>2</sub>-mediated eicosanoid production work through two distinct pathways. | en |
| dc.description.version | Published version | en |
| dc.format.extent | 22834 - 22847 page(s) | en |
| dc.identifier.doi | https://doi.org/10.1074/jbc.M701831200 | en |
| dc.identifier.eissn | 1083-351X | en |
| dc.identifier.issn | 0021-9258 | en |
| dc.identifier.issue | 31 | en |
| dc.identifier.uri | http://hdl.handle.net/10919/74311 | en |
| dc.identifier.volume | 282 | en |
| dc.rights | Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International | en |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | en |
| dc.title | TLR-4 and Sustained Calcium Agonists Synergistically Produce Eicosanoids Independent of Protein Synthesis in RAW264.7 Cells | en |
| dc.title.serial | Journal of Biological Chemistry | en |
| dc.type | Article - Refereed | en |
| pubs.organisational-group | /Virginia Tech | en |
| pubs.organisational-group | /Virginia Tech/All T&R Faculty | en |
| pubs.organisational-group | /Virginia Tech/Science | en |
| pubs.organisational-group | /Virginia Tech/Science/COS T&R Faculty | en |