Abstract:
:We report a new method for the detection of ligand-induced receptor internalization by fluorescence ratiometric imaging of pH in endosomes in combination with a recently developed posttranslational labeling system based on the formation of a heterodimeric coiled-coil structure. The N-terminus of the β2-adrenergic receptor expressed on the cell surface was doubly labeled with pH-sensitive fluorescein and pH-insensitive tetramethylrhodamine. A significant increase in the tetramethylrhodamine-to-fluorescein fluorescence intensity ratio was observed after incubation with agonists in a concentration-dependent manner. This simple and accurate method of detecting the agonistic activity of receptors will be useful for high-throughput screening of drug candidates.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Yano Y,Matsuzaki Kdoi
10.1016/j.febslet.2011.06.012subject
Has Abstractpub_date
2011-07-21 00:00:00pages
2385-8issue
14eissn
0014-5793issn
1873-3468pii
S0014-5793(11)00474-1journal_volume
585pub_type
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