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• W2044811761 abstract "1 [3H]-bradykinin was used to characterize the bradykinin receptors associated with canine cultured tracheal smooth muscle cells (TSMCs). Receptor binding assay showed that TSMCs had specific, saturable, high-affinity binding sites for [3H]-bradykinin. 2 The specific [3H]-bradykinin binding increased linearly with increasing cell concentrations. The equilibrium for association of [3H]-bradykinin with the bradykinin receptors was attained within 2 h at 4°C and 1 h at room temperature, respectively. 3 Analysis of binding isotherms yielded an apparent equilibrium dissociation constant (KD) of 2.5 ± 0.3 nm and a maximum receptor density (Bmax) of 25.1 ± 0.3 fmol mg−1 protein. The Hill coefficient for [3H]-bradykinin binding was 1.00 ± 0.02. The association (K1) and dissociation (K-1) rate constants were (8.67 ± 2.60) × 106 m−1 min−1 and 0.024 ± 0.005 min−1, respectively. KD, calculated from the ratio of K-1 and K1 was 2.8 ± 0.5 nm, a value close to that of KD calculated from Scatchard plots of binding isotherms. 4 The B1 receptor selective agonist, (des-Arg9-bradykinin, 0.1 nm–10 μm) and antagonist ([Leu8, des-Arg9]-bradykinin, 0.1 nm–10 μm) did not inhibit the [3H]-bradykinin binding to TSMCs, which excludes the presence of B1 receptors in canine TSMCs. 5 The specific binding of [3H]-bradykinin to canine TSMCs was inhibited by B2 receptor selective antagonists ([d-Arg0, Hyp3, Thi5, d-Tic7, Oic8]-bradykinin, Hoe 140, 0.1 nm–10 μm and [d-Arg0, Hyp3, Thi5,8, d-Phe7]-bradykinin, 0.1 nm–10 μm) and agonists (bradykinin and kallidin, 0.1 nm–10 μm) with a best fit by a one-binding site model. The order of potency for the inhibition of [3H]-bradykinin binding was kallidin = bradykinin = Hoe 140>[d-Arg0, Hyp3, Thi5,8, d-Phe7]-bradykinin. 6 Preincubation of TSMCs with forskolin for 24 h led to an up-regulation of B2 receptors, increasing in Bmax from 25.1 ± 0.3 to 218 ± 24 fmol mg−1 protein without changing the KD values. [3H]-bradykinin binding to TSMCs was inhibited by the B2 receptor selective antagonists and agonists, but not by the B1 receptor selective reagents. The up-regulation of the B2 receptor by forskolin was mediated through protein synthesis, since cycloheximide blocked this response. 7 It is concluded that the pharmacological characteristics of the bradykinin receptors in canine cultured TSMCs are primarily of the B2 receptor subtype." @default.
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• W2044811761 date "1995-01-01" @default.
• W2044811761 modified "2023-10-16" @default.
• W2044811761 title "Pharmacological characterization of bradykinin receptors in canine cultured tracheal smooth muscle cells" @default.
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• W2044811761 doi "https://doi.org/10.1111/j.1476-5381.1995.tb14906.x" @default.
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