A pH-sensitive fluor, CypHer 5, used to monitor agonist-induced G protein-coupled receptor internalization in live cells

A pH-sensitive fluor, CypHer 5, used to monitor agonist-induced G protein-coupled receptor internalization in live cells

G protein-coupled receptors (GPCRs) are the largest family of proteins involved in transmembrane signal transduction and are actively studied because of their suitability as therapeutic small-molecule drug targets. Agonist activation of GPCRs almost invariably results in the receptor being desensiti...

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Veröffentlicht in:BioTechniques. - 1988. - 33(2002), 5 vom: 21. Nov., Seite 1152-4, 1156-7
1. Verfasser: Adie, E J (VerfasserIn)
Weitere Verfasser: Kalinka, S, Smith, L, Francis, M J, Marenghi, A, Cooper, M E, Briggs, M, Michael, N P, Milligan, G, Game, S
Format: Aufsatz
Sprache:English
Veröffentlicht: 2002
Zugriff auf das übergeordnete Werk:BioTechniques
Schlagworte:Evaluation Study Journal Article Antibodies, Anti-Idiotypic Carbocyanines CypHer 5 NHS ester Epitopes Fluorescent Dyes G protein, vesicular stomatitis virus Luminescent Proteins Membrane Glycoproteins mehr... PTGIR protein, human Ptgir protein, mouse Receptors, Epoprostenol Receptors, Opioid, delta Receptors, Prostaglandin Receptors, Thyrotropin-Releasing Hormone Recombinant Fusion Proteins Viral Envelope Proteins anti-IgG Green Fluorescent Proteins 147336-22-9 Enkephalin, Leucine-2-Alanine 63631-40-3 Thyrotropin 9002-71-5 GTP-Binding Proteins EC 3.6.1.- Iloprost JED5K35YGL
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100 1 |a Adie, E J  |e verfasserin  |4 aut 
245 1 2 |a A pH-sensitive fluor, CypHer 5, used to monitor agonist-induced G protein-coupled receptor internalization in live cells 
264 1 |c 2002 
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500 |a Date Completed 17.06.2003 
500 |a Date Revised 18.03.2022 
500 |a published: Print 
500 |a Citation Status MEDLINE 
520 |a G protein-coupled receptors (GPCRs) are the largest family of proteins involved in transmembrane signal transduction and are actively studied because of their suitability as therapeutic small-molecule drug targets. Agonist activation of GPCRs almost invariably results in the receptor being desensitized. One of the key events in receptor desensitization is the sequestration of the receptor from the cell surface into acidic intracellular endosomes. Therefore, a convenient, generic, and noninvasive monitor of this process is desirable. A novel, pH-sensitive, red-excited fluorescent dye, CypHer 5, was synthesized. This dye is non-fluorescent at neutral pH and is fluorescent at acidic pH. Anti-epitope antibodies labeled with this dye were internalized in an agonist concentration- and time-dependent manner, following binding on live cells to a range of GPCRs that had been modified to incorporate the epitope tags in their extracellular N-terminal domain. This resulted in a large signal increase over background. When protonated, the red fluorescence of CypHer 5 provides a generic reagent suitable for monitoring the internalization of GPCRs into acidic vesicles. This approach should be amenable to the study of many other classes of cell surface receptors that also internalize following stimulation 
650 4 |a Evaluation Study 
650 4 |a Journal Article 
650 7 |a Antibodies, Anti-Idiotypic  |2 NLM 
650 7 |a Carbocyanines  |2 NLM 
650 7 |a CypHer 5 NHS ester  |2 NLM 
650 7 |a Epitopes  |2 NLM 
650 7 |a Fluorescent Dyes  |2 NLM 
650 7 |a G protein, vesicular stomatitis virus  |2 NLM 
650 7 |a Luminescent Proteins  |2 NLM 
650 7 |a Membrane Glycoproteins  |2 NLM 
650 7 |a PTGIR protein, human  |2 NLM 
650 7 |a Ptgir protein, mouse  |2 NLM 
650 7 |a Receptors, Epoprostenol  |2 NLM 
650 7 |a Receptors, Opioid, delta  |2 NLM 
650 7 |a Receptors, Prostaglandin  |2 NLM 
650 7 |a Receptors, Thyrotropin-Releasing Hormone  |2 NLM 
650 7 |a Recombinant Fusion Proteins  |2 NLM 
650 7 |a Viral Envelope Proteins  |2 NLM 
650 7 |a anti-IgG  |2 NLM 
650 7 |a Green Fluorescent Proteins  |2 NLM 
650 7 |a 147336-22-9  |2 NLM 
650 7 |a Enkephalin, Leucine-2-Alanine  |2 NLM 
650 7 |a 63631-40-3  |2 NLM 
650 7 |a Thyrotropin  |2 NLM 
650 7 |a 9002-71-5  |2 NLM 
650 7 |a GTP-Binding Proteins  |2 NLM 
650 7 |a EC 3.6.1.-  |2 NLM 
650 7 |a Iloprost  |2 NLM 
650 7 |a JED5K35YGL  |2 NLM 
700 1 |a Kalinka, S  |e verfasserin  |4 aut 
700 1 |a Smith, L  |e verfasserin  |4 aut 
700 1 |a Francis, M J  |e verfasserin  |4 aut 
700 1 |a Marenghi, A  |e verfasserin  |4 aut 
700 1 |a Cooper, M E  |e verfasserin  |4 aut 
700 1 |a Briggs, M  |e verfasserin  |4 aut 
700 1 |a Michael, N P  |e verfasserin  |4 aut 
700 1 |a Milligan, G  |e verfasserin  |4 aut 
700 1 |a Game, S  |e verfasserin  |4 aut 
773 0 8 |i Enthalten in  |t BioTechniques  |d 1988  |g 33(2002), 5 vom: 21. Nov., Seite 1152-4, 1156-7  |w (DE-627)NLM012627046  |x 1940-9818  |7 nnns 
773 1 8 |g volume:33  |g year:2002  |g number:5  |g day:21  |g month:11  |g pages:1152-4, 1156-7 
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