Differential Allosteric Modulation by Amiloride Analogues of Agonist and Antagonist Binding at A(1) and A(3) Adenosine Receptors

Overview

Journal Biochem Pharmacol

Specialties Biochemistry
Pharmacology

Date 2003 Feb 5

PMID 12566079

Citations 23

Authors

Zhan-Guo Gao

Neli Melman

Andreas Erdmann

Seong Gon Kim

Christa E Muller

Adriaan P IJzerman

Kenneth A Jacobson

Affiliations

Soon will be listed here.

Abstract

The diuretic drug amiloride and its analogues were found previously to be allosteric modulators of antagonist binding to A(2A) adenosine receptors. In this study, the possibility of the allosteric modulation by amiloride analogues of antagonist binding at A(1) and A(3) receptors, as well as agonist binding at A(1), A(2A), and A(3) receptors, was explored. Amiloride analogues increased the dissociation rates of two antagonist radioligands, [3H]8-cyclopentyl-1,3-dipropylxanthine ([3H]DPCPX) and [3H]8-ethyl-4-methyl-2-phenyl-(8R)-4,5,7,8-tetrahydro-1H-imidazo[2,1-i]purin-5-one ([3H]PSB-11), from A(1) and A(3) receptors, respectively. Amiloride and 5-(N,N-dimethyl)amiloride (DMA) were more potent at A(1) receptors than at A(3) receptors, while 5-(N,N-hexamethylene)amiloride (HMA) was more potent at A(3) receptors. Thus, amiloride analogues are allosteric inhibitors of antagonist binding at A(1), A(2A), and A(3) adenosine receptor subtypes. In contrast to their effects on antagonist-occupied receptors, amiloride analogues did not affect the dissociation rates of the A(1) agonist [3H]N(6)-[(R)-phenylisopropyl]adenosine ([3H]R-PIA) from A(1) receptors or the A(2A) agonist [3H]2-[p-(2-carboxyethyl)phenyl-ethylamino]-5'-N-ethylcarboxamidoadenosine ([3H]CGS21680) from A(2A) receptors. The dissociation rate of the A(3) agonist radioligand [125I]N(6)-(4-amino-3-iodobenzyl)adenosine-5'-N-methyluronamide ([125I]I-AB-MECA) from A(3) receptors was decreased significantly by amiloride analogues. The binding modes of amiloride analogues at agonist-occupied and antagonist-occupied receptors differed markedly, which was demonstrated in all three subtypes of adenosine receptors tested in this study. The effects of the amiloride analogues on the action of the A(3) receptor agonist were explored further using a cyclic AMP functional assay in intact CHO cells expressing the human A(3) receptor. Both binding and functional assays support the allosteric interactions of amiloride analogues with A(3) receptors.

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References

Howard M, Hughes R, Motulsky H, Mullen M, Insel P . Interactions of amiloride with alpha- and beta-adrenergic receptors: amiloride reveals an allosteric site on alpha 2-adrenergic receptors. Mol Pharmacol. 1987; 32(1):53-8. View

Gao Z, van Muijlwijk-Koezen J, Chen A, Muller C, IJzerman A, Jacobson K . Allosteric modulation of A(3) adenosine receptors by a series of 3-(2-pyridinyl)isoquinoline derivatives. Mol Pharmacol. 2001; 60(5):1057-63. PMC: 3953614. View

Nordstedt C, Fredholm B . A modification of a protein-binding method for rapid quantification of cAMP in cell-culture supernatants and body fluid. Anal Biochem. 1990; 189(2):231-4. DOI: 10.1016/0003-2697(90)90113-n. View

Gao Z, Kim S, Soltysiak K, Melman N, IJzerman A, Jacobson K . Selective allosteric enhancement of agonist binding and function at human A3 adenosine receptors by a series of imidazoquinoline derivatives. Mol Pharmacol. 2002; 62(1):81-9. PMC: 3953617. DOI: 10.1124/mol.62.1.81. View

Urwyler S, Mosbacher J, Lingenhoehl K, Heid J, Hofstetter K, Froestl W . Positive allosteric modulation of native and recombinant gamma-aminobutyric acid(B) receptors by 2,6-Di-tert-butyl-4-(3-hydroxy-2,2-dimethyl-propyl)-phenol (CGP7930) and its aldehyde analog CGP13501. Mol Pharmacol. 2001; 60(5):963-71. View

Bhattacharya S, Linden J . Effects of long-term treatment with the allosteric enhancer, PD81,723, on Chinese hamster ovary cells expressing recombinant human A1 adenosine receptors. Mol Pharmacol. 1996; 50(1):104-11. View

Horstman D, Brandon S, Wilson A, Guyer C, Cragoe Jr E, Limbird L . An aspartate conserved among G-protein receptors confers allosteric regulation of alpha 2-adrenergic receptors by sodium. J Biol Chem. 1990; 265(35):21590-5. View

Jacobson K, Park K, Jiang J, Kim Y, Olah M, Stiles G . Pharmacological characterization of novel A3 adenosine receptor-selective antagonists. Neuropharmacology. 1997; 36(9):1157-65. PMC: 3433714. DOI: 10.1016/s0028-3908(97)00104-4. View

Kobilka B . Allosteric activation of the CaR by L-amino acids. Proc Natl Acad Sci U S A. 2000; 97(9):4419-20. PMC: 34312. DOI: 10.1073/pnas.97.9.4419. View

10.

Olah M, Stiles G . The role of receptor structure in determining adenosine receptor activity. Pharmacol Ther. 2000; 85(2):55-75. DOI: 10.1016/s0163-7258(99)00051-0. View

11.

Kleyman T, Cragoe Jr E . Cation transport probes: the amiloride series. Methods Enzymol. 1990; 191:739-55. DOI: 10.1016/0076-6879(90)91045-8. View

12.

Leppik R, Lazareno S, Mynett A, Birdsall N . Characterization of the allosteric interactions between antagonists and amiloride analogues at the human alpha2A-adrenergic receptor. Mol Pharmacol. 1998; 53(5):916-25. View

13.

ARUNLAKSHANA O, SCHILD H . Some quantitative uses of drug antagonists. Br J Pharmacol Chemother. 1959; 14(1):48-58. PMC: 1481829. DOI: 10.1111/j.1476-5381.1959.tb00928.x. View

14.

Birdsall N, Cohen F, Lazareno S, Matsui H . Allosteric regulation of G-protein-linked receptors. Biochem Soc Trans. 1995; 23(1):108-11. DOI: 10.1042/bst0230108. View

15.

Hoare S, Strange P . Regulation of D2 dopamine receptors by amiloride and amiloride analogs. Mol Pharmacol. 1996; 50(5):1295-308. View

16.

Bruns R, Fergus J . Allosteric enhancement of adenosine A1 receptor binding and function by 2-amino-3-benzoylthiophenes. Mol Pharmacol. 1990; 38(6):939-49. View

17.

Neve K, Cumbay M, Thompson K, Yang R, Buck D, Watts V . Modeling and mutational analysis of a putative sodium-binding pocket on the dopamine D2 receptor. Mol Pharmacol. 2001; 60(2):373-81. DOI: 10.1124/mol.60.2.373. View

18.

Litschig S, Gasparini F, Rueegg D, Stoehr N, Flor P, Vranesic I . CPCCOEt, a noncompetitive metabotropic glutamate receptor 1 antagonist, inhibits receptor signaling without affecting glutamate binding. Mol Pharmacol. 1999; 55(3):453-61. View

19.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

20.

Gao Z, IJzerman A . Allosteric modulation of A(2A) adenosine receptors by amiloride analogues and sodium ions. Biochem Pharmacol. 2000; 60(5):669-76. DOI: 10.1016/s0006-2952(00)00360-9. View