Fluorescence ratiometric detection of ligand-induced receptor internalization using extracellular coiled-coil tag-probe labeling.

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 Lett

journal_title

FEBS letters

authors

Yano Y,Matsuzaki K

doi

10.1016/j.febslet.2011.06.012

subject

Has Abstract

pub_date

2011-07-21 00:00:00

pages

2385-8

issue

14

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(11)00474-1

journal_volume

585

pub_type

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