Assessing activation of the human neuropeptide FF2 receptor with a non-radioactive GTP binding assay.
We have evaluated a novel, time-resolved fluorometric GTP binding assay for its suitability for functional screening of neuropeptide FF (NPFF) receptor ligands. Our results suggest that this assay, which relies on the use of a europium-labeled GTP analogue, Eu-GTP, provides a powerful alternative to the [35S]guanosine-5'-O-(3-thio)triphosphate binding assay for assessing the functional properties of NPFF analogs. Further, we demonstrate that the tetrapeptide PMRF-NH2 exhibited high agonist potency at the NPFF2 receptor, and that the efficacies of this peptide and another shortened NPFF analog were greater than that of NPFF.
Engström
Mia
M
Juvantia Pharma Ltd., Lemminkäisenkatu 5, Pharmacity, FIN-20520 Turku, Finland. mia.engstrom@juvantia.com
Närvänen
Ale
A
Savola
Juha-Matti
JM
Wurster
Siegfried
S
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D005063:Europium; D005470:Fluorometry; D006160:Guanosine Triphosphate / Q000378:metabolism*; D018013:Receptors, Neuropeptide / Q000819:agonists / Q000378:metabolism*; D013329:Structure-Activity Relationship
D018013:Receptors, Neuropeptide; C083929:neuropeptide FF receptor; D005063:Europium; D006160:Guanosine Triphosphate