Biology Reference
In-Depth Information
TAS2R CoIP studies
in vivo
. However, the availability of adequate amounts of tissue
still provides a challenge.
BRET experiments on the other hand could be useful to analyze many different
receptor variants or mutants for their ability to form oligomers, for example, regard-
ing the functionality of the heteromeric sweet taste receptor or, for example, regard-
ing the identification of the domain(s) involved in and necessary for oligomerization
of TAS1Rs and TAS2Rs, respectively.
SUMMARY
We have here provided a detailed protocol for the investigation of the oligomeriza-
tion of both TAS1R and TAS2R taste receptor families. Starting with the easy and
straightforward generation of receptor constructs needed for the following methods,
we also provide a protocol for quick analysis of expression of these constructs in
mammalian cell lines. We have then detailed CoIP experiments for the biochemical
analysis of protein-protein interactions, incorporating important controls into our
protocol. Further, we have also explained the use of BRET for the investigation
of receptor oligomerization in living cells and for a high throughput. The protocols
provided here should also be easily transferable to studies on oligomerization of
other receptors, for example, other rhodopsin-like GPCRs or other class C GPCRs.
Acknowledgments
This work was supported by the German Research Foundation (DFG, Me 1024/2-1, 2-2).
References
