| First Author | Kobe F | Year | 2008 |
| Journal | Biochim Biophys Acta | Volume | 1783 |
| Issue | 8 | Pages | 1503-16 |
| PubMed ID | 18381076 | Mgi Jnum | J:136992 |
| Mgi Id | MGI:3797467 | Doi | 10.1016/j.bbamcr.2008.02.021 |
| Citation | Kobe F, et al. (2008) Stimulation- and palmitoylation-dependent changes in oligomeric conformation of serotonin 5-HT1A receptorsi. Biochim Biophys Acta 1783(8):1503-16 |
| abstractText | In the present study we analyzed the oligomerization state of the serotonin 5-HT1A receptor and studied oligomerization dynamics in living cells. We also investigated the role of receptor palmitoylation in this process. Biochemical analysis performed in neuroblastoma N1E-115 cells demonstrated that both palmitoylated and non-palmitoylated 5-HT1A receptors form homo-oligomers and that the prevalent receptor species at the plasma membrane are dimers. A combination of an acceptor-photobleaching FRET approach with fluorescence lifetime measurements verified the interaction of CFP- and YFP-labeled wild-type as well as acylation-deficient 5-HT1A receptors at the plasma membrane of living cells. Using a novel FRET technique based on the spectral analysis we also confirmed the specific nature of receptor oligomerization. The analysis of oligomerization dynamics revealed that apparent FRET efficiency measured for wild-type oligomers significantly decreased in response to agonist stimulation, and our combined results suggest that this decrease was mediated by accumulation of FRET-negative complexes rather than by dissociation of oligomers to monomers. In contrast, the agonist-mediated decrease of FRET signal was completely abolished in oligomers composed by non-palmitoylated receptor mutants, demonstrating the importance of palmitoylation in modulation of the structure of oligomers. |
