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Pharmacological characterization of unique prazosin-binding sites in human kidney

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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.

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Acknowledgements

This work was supported in part by grant from the Smoking Research Foundation of Japan and Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, Fukui Medical University, 910–1193, Matsuoka, Fukui, Japan

    Yasuko Hiraoka, Takanobu Taniguchi, Takashi Tanaka & Ikunobu Muramatsu

  2. Department of Urology, School of Medicine, Fukui Medical University, 910–1193, Matsuoka, Fukui, Japan

    Kenichiro Okada & Hiroshi Kanamaru

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  1. Yasuko Hiraoka
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  2. Takanobu Taniguchi
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  3. Takashi Tanaka
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Correspondence to Ikunobu Muramatsu.

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Hiraoka, Y., Taniguchi, T., Tanaka, T. et al. Pharmacological characterization of unique prazosin-binding sites in human kidney. Naunyn-Schmiedeberg's Arch Pharmacol 368, 49–56 (2003). https://doi.org/10.1007/s00210-003-0764-x

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