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Pharmacological characterization of a novel non-AT1, non-AT2 angiotensin binding site identified as neurolysin

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Abstract

The discovery of a novel non-AT1, non-AT2 binding site for angiotensins in the rodent brain and testis that is unmasked by the organomercurial compound para-chloromercuribenzoic acid (PCMB) has catalyzed efforts to purify and characterize this protein. We recently reported that this protein is neurolysin and now report upon the specificity of this binding site for various neuropeptides. Competition binding assays in rat brain and testis used 125I-Sar1, Ile8 angiotensin II (Ang II) as the radioligand in the presence of saturating concentrations of AT1 and AT2 receptor antagonists and 100 μM parachloromercuribenzoate. Primary screening of 36 peptides and other compounds at 10 μM concentration revealed seven peptides that inhibited specific binding >50 %: ghrelin, Tyr1 S36057 (a melanin-concentrating hormone receptor ligand), orphanin FQ and its congeners (Tyr1 and Tyr14), Dynorphin A (1–8), and Ang (1–9). The selective neurolysin inhibitor Proline–Isoleucine dipeptide was inactive at 1 mM. These results suggest that the ability of PCMB to unmask high affinity binding of Ang II to neurolysin is a pharmacological effect and that neurolysin may significantly affect the activity of the renin-angiotensin system.

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Acknowledgments

The authors thank Lancya Lansdowne for technical assistance, Drs. Gerhard Munskie and Nour-Eddine Rhaleb for providing angiotensin system-related peptides. We thank Drs. Vardan Karamyan and Michelle Clark and Mr. Eduardo Carrera for their critical comments and review of this manuscript. We thank Dr. Colin Sumners for providing the rat tissues used for these studies. This work was supported by NIH NHLBI HL-096357.

Conflict of interest

Neither Dr. Robert C. Speth, Kira L. Santos nor Jamala D. Swindle have any conflicts of interests or competing interests with respect to the publication of our manuscript: “Pharmacological characterization of a novel non-AT1, non-AT2 angiotensin-binding site identified as neurolysin” which we have submitted to Endocrine for consideration for publication.

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Authors and Affiliations

  1. Farquhar College of Arts and Sciences, Nova Southeastern University, Fort Lauderdale, FL, 33314, USA

    Jamala D. Swindle

  2. Department of Pharmaceutical Sciences, College of Pharmacy, Nova Southeastern University, 3200 S University Drive, Fort Lauderdale, FL, 33328, USA

    Jamala D. Swindle, Kira L. Santos & Robert C. Speth

  3. Department of Physiology and Functional Genomics, College of Medicine, University of Florida, Gainesville, FL, 32610, USA

    Robert C. Speth

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  1. Jamala D. Swindle
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Correspondence to Robert C. Speth.

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Swindle, J.D., Santos, K.L. & Speth, R.C. Pharmacological characterization of a novel non-AT1, non-AT2 angiotensin binding site identified as neurolysin. Endocrine 44, 525–531 (2013). https://doi.org/10.1007/s12020-013-9898-x

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