Inhibition of enzymatic degradation of angiotensin II in membrane binding assays: utility of 1,10-phenanthroline

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Abstract
Studies of 125I-labelled angiotensin II binding to membranes from bovine uterus and aorta are rendered impossible because enzymatic degradation of 90% of the added ligand occurs within 30 min at 23 °C. High pressure liquid chromatographic analysis has demonstrated that a single iodinated product, which is more hydrophilic than intact angiotensin II, is produced by enzymatic cleavage. Investigation of a series of enzyme inhibitors has shown that the carboxypeptidase inhibitor 1,10-phenanthroline completely inhibits degradation of angiotensin II without disrupting ligand binding.Key words: angiotensin II, binding, degradation, bovine uterine membrane, bovine aortic membrane, 1,10-phenanthroline inhibitor.