{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T05:38:23Z","timestamp":1784266703641,"version":"3.55.0"},"posted":{"date-parts":[[2024,4,12]]},"group-title":"Pharmacology and Toxicology","reference-count":48,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,4,18]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>The renin-angiotensin system involves many more enzymes, receptors and biologically active peptides than originally thought. With this study, we investigated whether angiotensin-(1-5) [Ang-(1-5)], a 5-amino acid fragment of angiotensin II, has biological activity, and through which receptor it elicits effects.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    The effect of Ang-(1-5) (1\u00b5M) on nitric oxide release was measured by DAF-FM staining in human aortic endothelial cells (HAEC), or Chinese Hamster Ovary (CHO) cells stably transfected with the angiotensin AT\n                    <jats:sub>2<\/jats:sub>\n                    -receptor (AT\n                    <jats:sub>2<\/jats:sub>\n                    R) or the receptor Mas. A potential vasodilatory effect of Ang-(1-5) was tested in mouse mesenteric and human renal arteries by wire myography; the effect on blood pressure was evaluated in normotensive C57BL\/6 mice by Millar catheter. These experiments were performed in the presence or absence of a range of antagonists or inhibitors or in AT\n                    <jats:sub>2<\/jats:sub>\n                    R-knockout mice. Binding of Ang-(1-5) to the AT\n                    <jats:sub>2<\/jats:sub>\n                    R was confirmed and the preferred conformations determined by\n                    <jats:italic>in silico<\/jats:italic>\n                    docking simulations. The signaling network of Ang-(1-5) was mapped by quantitative phosphoproteomics.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Key findings included: (1) Ang-(1-5) induced activation of eNOS by changes in phosphorylation at\n                    <jats:sup>Ser1177<\/jats:sup>\n                    eNOS and\n                    <jats:sup>Tyr657<\/jats:sup>\n                    eNOS and thereby (2) increased NO release from HAEC and AT\n                    <jats:sub>2<\/jats:sub>\n                    R-transfected CHO cells, but not from Mas-transfected or non-transfected CHO cells. (3) Ang-(1-5) induced relaxation of preconstricted mouse mesenteric and human renal arteries and (4) lowered blood pressure in normotensive mice \u2013 effects which were respectively absent in arteries from AT\n                    <jats:sub>2<\/jats:sub>\n                    R-KO or in PD123319-treated mice and which were more potent than effects of the established AT\n                    <jats:sub>2<\/jats:sub>\n                    R-agonist C21. (5) According to\n                    <jats:italic>in silico<\/jats:italic>\n                    modelling, Ang-(1-5) binds to the AT\n                    <jats:sub>2<\/jats:sub>\n                    R in two preferred conformations, one differing substantially from where the first five amino acids within angiotensin II bind to the AT\n                    <jats:sub>2<\/jats:sub>\n                    R. (6) Ang-(1-5) modifies signaling pathways in a protective RAS-typical way and with relevance for endothelial cell physiology and disease.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>\n                    Ang-(1-5) is a potent, endogenous AT\n                    <jats:sub>2<\/jats:sub>\n                    R-agonist.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2024.04.05.588367","type":"posted-content","created":{"date-parts":[[2024,4,12]],"date-time":"2024-04-12T16:10:16Z","timestamp":1712938216000},"source":"Crossref","is-referenced-by-count":7,"title":["Angiotensin-(1-5) is a Potent Endogenous Angiotensin AT\n                  <sub>2<\/sub>\n                  -Receptor Agonist"],"prefix":"10.64898","author":[{"given":"Igor M.","family":"Souza-Silva","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"A. 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