Development of a binding assay for the B1 receptors for kinins

Agonist-Induced Translocation of the Kinin B1Receptor to Caveolae-Related Rafts

“…4. The radioligand binding was essentially complete after 60 min at 37°C, but was extremely slow at 0°(incomplete after 2 h), as reported for the rabbit wild-type B 1 R expressed by smooth muscle cells (Levesque et al, 1995). After ligand association at 37°C, some wells were washed three times with PBS at time 60 min, and further incubated in the azide-free binding medium for 4 h either at molpharm.aspetjournals.org 37 or 0°C.…”

“…An additional common feature of the wild-type rabbit B 1 R and B 1 R-YFP is the exceptionally slow radioligand association at 0°C in intact cells ( Fig. 4; Levesque et al, 1995). A further functional cellular response mediated by the fusion protein is agonistinduced redistribution of B 1 R-YFP, a temperature-dependent effect occurring at low concentrations of Lys-desArg 9 -BK (1 nM; Fig.…”

Non‐competitive pharmacological antagonism at the rabbit B1 receptor

“…We conducted our radioligand binding assay at 378C in the presence of sodium azide; these conditions allow reaching an e ective equilibrium without ligand internalization in this system (Levesque et al, 1995a). The observed control K D values were in the same range than those obtained using rabbit smooth muscle cells and the same tritiated ligand (Schneck et al, 1994;Galizzi et al, 1994).…”

“…A binding assay to rabbit B 1 receptors was conducted as described (Levesque et al, 1995a), except for the identity of the ligand which was [ 3 H]-Lys-des-Arg 9 -BK ([ 3 H]-des-Arg 10 -kallidin, NEN Biosciences, 105 Ci mmol 71 ). Brie¯y, COS-1 cells were seeded at a high density in 24-well plates (culture medium DMEM supplemented with 10% foetal bovine serum and antibiotics, Gibco).…”

N-terminal extended conjugates of the agonists and antagonists of both bradykinin receptor subtypes: Structure–activity relationship, cell imaging using ligands conjugated with fluorophores and prospect for functionally active cargoes

“…Gly-Lys--aminocaproyl-Lys-des-Arg 9 -BK [22], successfully used in tumor autoradiography [25], or of the highly potent antagonist B-10324 (2,3,4,5,6-pentafluorocinnamoyl-B-9958) [16].…”