Novel high molecular weight albumin-conjugated angiotensin II activates β-arrestin and G-protein pathways

Purpose: To study the ability of a novel bovine serum albumin-angiotensin II (BSA-Ang II) conjugate to effect responses of the AT₁ angiotensin II receptor subtype mediated by the G-protein-coupled and the beta-arrestin pathways. Methods: Angiotensin II (Ang II) was conjugated with bovine serum albumin and compared with Ang II for competition binding to AT₁ receptors, to stimulate aldosterone release from adrenocortical cells, to promote beta-arrestin binding to AT₁ receptors, to promote calcium mobilization, and stimulate drinking of water and saline by rats. Results: The BSA-Ang II conjugate was less potent competing for AT₁R binding, but was equally efficacious at stimulating aldosterone release from H295R adrenocortical cells. Both BSA-Ang II and Ang II stimulated calcium mobilization and beta-arrestin binding to AT₁ receptors. BSA-Ang II and Ang II stimulated water appetite equivalently but BSA-Ang II stimulated saline appetite more than Ang II. Both BSA-Ang II and Ang II were considerably more potent at causing calcium mobilization than β-arrestin binding. Conclusions: Addition of a high molecular weight molecule to Ang II reduced its AT₁ receptor binding affinity, but did not significantly alter stimulation of aldosterone release or water consumption. The BSA-Ang II conjugate caused a greater saline appetite than Ang II suggesting that it may be a more efficacious agonist of this beta-arrestin-mediated response than Ang II. The higher potency calcium signaling response suggests that the G-protein-coupled responses predominate at physiological concentrations of Ang II, while the beta-arrestin response requires pathophysiological or pharmacological concentrations of Ang II to occur.