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M W Beukers

Publications and source records attributed to M W Beukers.

20 records · Page 2Linked to original sources
PubMed1990

Interaction of amiloride and its analogues with adenosine A1 receptors in calf brain.

Amiloride, a potassium sparing diuretic, is known to interact with a number of ion transport systems, receptors and enzymes. Here, we report on the interaction between this drug and the adenosine A1 receptor as present in calf brain membranes. Adenosine A1 receptors are characterized by a subnanomolar affinity for the antagonists [3H]8-cyclopentyl-1,3-dipropylxanthine ([3H]DPCPX) and the agonist [3H]N6-R-1-phenyl-2-propyladenosine ([3H]PIA). Amiloride displaces both agonist and antagonist binding with a Ki value in the low micromolar range. This inhibition is counteracted by NaCl and protons, in contrast to the binding of [3H]PIA and [3H]DPCPX. The results suggest that amiloride interacts with the adenosine A1 receptor at a site distinct from the ligand binding site. In order to elucidate the role of one of the ion transport systems known to be inhibited by amiloride, eight amiloride analogues with different sensitivities for these systems were tested. The potency and order of potency of these compounds towards adenosine A1 receptors excludes the involvement of the epithelial Na+ channel, Na+/H+ exchanger or Na+/Ca2+ exchanger.

Adenosine↗
PubMed1990

Chemical modification of adenosine A1 receptors. Implications for the interaction with R-PIA, DPCPX and amiloride.

Amiloride, a potassium sparing diuretic, inhibits the specific binding of [3H]8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and [3H]N6-R-1-phenyl-2-propyladenosine (PIA) to adenosine A1 receptors in calf brain. This interaction is different from the agonist-receptor or the antagonist-receptor interaction as Na+ and H+ counteract the inhibitory effect of amiloride whereas these ions hardly affect the binding of the classic A1 receptor ligands. In the present study, the effects of protein modifiers on the equilibrium inhibition constant of amiloride are compared with effects of these reagents on the affinities of DPCPX and PIA. It is demonstrated that the affinities of amiloride and [3H]DPCPX are changed after treatment with a carboxyl-modifying reagent but unaffected by modification of histidyl, arginyl and cystein residues. The maximal binding capacity of [3H]DPCPX is enhanced by sulfhydryl modification, whereas the number of [3H]DPCPX binding sites is reduced by treatment with a histidine-modifying reagent. The histidyl residues of the [3H]DPCPX binding site can be partially protected against modification by 300 microM amiloride, present during treatment of the membranes. An equivalent concentration of 8-phenyltheophylline results in complete protection. The apparent affinity of PIA is altered by modification of histidyl, carboxyl, arginyl and cystein residues. In the latter two cases, uncoupling of the G protein seems to be the major reason for the decrease in affinity of PIA. The results suggest that amiloride is an A1 antagonist with binding characteristics that differ from the classic A1 antagonists such as DPCPX.

Adenosine↗