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- Title
Pharmacological characterization of unique prazosin-binding sites in human kidney.
- Authors
Hiraoka, Yasuko; Taniguchi, Takanobu; Tanaka, Takashi; Okada, Kenichiro; Kanamaru, Hiroshi; Muramatsu, Ikunobu
- Abstract
In human kidney, we found unique prazosin-binding sites that were insensitive to phentolamine and were thus unlikely to be α1-adrenoceptors. As the binding of [3H]prazosin to phentolamine-insensitive sites was prevented by 100 μM guanabenz, the insensitive sites were evaluated by subtracting [3H]prazosin binding in the presence of 100 μM guanabenz from that in the presence of 10 μM phentolamine. [3H]Prazosin bound to the phentolamine-insensitive sites monophasically with a high affinity (pKd; 9.1±0.08, n=8), and the Bmax value (814±204 fmol mg-1 protein, n=8) was more than ten times that of the phentolamine-sensitive α1-adrenoceptor (pKd=9.9±0.13, Bmax=66±23 fmol mg-1 protein, n=7). The phentolamine-insensitive sites in human kidney were highly sensitive to other quinazoline derivatives such as terazosin and doxazosin. However, other α1-adrenoceptor antagonists (tamsulosin, WB4101 and corynanthine) did not inhibit the binding at a range of concentrations that generally exhibit α1-adrenoceptor antagonism, and noradrenaline, rauwolscine and propranolol were without effect on the [3H]prazosin binding. On the other hand, ligands for the renal Na+-transporter (amiloride and triamterene) and for imidazoline recognition sites (guanabenz, guanfacine and agmatine) displaced the binding of [3H]prazosin to phentolamine-insensitive sites at micromolar concentrations. Photoaffinity labeling with [125I]iodoarylazidoprazosin showed phentolamine-insensitive labeling at around 100 kDa, a molecular size larger than that of human α1a- and α1b-adrenoceptors expressed in 293 cells (50–60 and 70–80 kDa, respectively) on electrophoresis. In contrast, there was no detectable phentolamine-insensitive binding site but were phentolamine-sensitive α1-adrenoceptors in human liver (pKd=10.0±0.06, Bmax=44±6 fmol mg–1 protein, n=3). Phentolamine-insensitive prazosin binding sites were also detected in rabbit kidney (approximately 50% of specific binding sites) but were minor in rat kidney (less than 20%). In conclusion, there are unique prazosin-binding sites in human kidney, the pharmacological profiles of which were distinct from those of known adrenoceptors.
- Subjects
KIDNEYS; PRAZOSIN; PHENTOLAMINE; PHOTOAFFINITY labeling; ADRENERGIC receptors; ELECTROPHORESIS
- Publication
Naunyn-Schmiedeberg's Archives of Pharmacology, 2003, Vol 368, Issue 1, p49
- ISSN
0028-1298
- Publication type
Article
- DOI
10.1007/s00210-003-0764-x