Human brain contains a novel non-AT1, non-AT2 binding site for active angiotensin peptides

AimsTo determine whether the novel non-AT1, non-AT2 binding site for angiotensins recently discovered in rodent brains occurs in the human brain.Main methodsRadioligand binding assays of 125I-sarcosine1, isoleucine8 angiotensin II binding were carried out in homogenates of the rostral pole of the temporal cortex of human brains containing 0.3 mM parachloromercuribenzoate (PCMB), 10 µM losartan to saturate AT1 receptors, 10 µM PD123319 to saturate AT2 receptors, with or without 10 µM angiotensin II to define specific binding. Competition binding assays employed a variety of angiotensin peptides, specific angiotensin receptor antagonists, several neuropeptides and an endopeptidase inhibitor to determine pharmacological specificity for this binding site.Key findingsThe novel non-AT1, non-AT2 binding site was present in similar amounts in female and male brains: Bmax 1.77 ± 0.16 and 1.52 ± 0.17 fmol/mg initial wet weight in female and male brains, respectively. The KD values, 1.79 ± 0.09 nM for females, and 1.53 ± 0.06 nM for males were also similar. The binding site shows pharmacological specificity similar to that in rodent brains: sarcosine1, isoleucine8 angiotensin II > angiotensin III > angiotensin II > angiotensin I ’ angiotensin IV > angiotensin 1–7. Shorter angiotensin fragments and non-angiotensin peptides showed low affinity for this binding site.SignificanceThe presence in human brain of this novel non-AT1, non-AT2 binding site supports the concept that this binding site is an important component of the brain angiotensin system. The functional significance of this binding site, either as a novel angiotensin receptor or a highly specific angiotensinase remains to be determined.