Fluorescently labeled adrenomedullin allows real-time monitoring of adrenomedullin receptor trafficking in living cells

Journal of Peptide Science : an Official Publication of the European Peptide Society
Ria SchönauerAnnette G Beck-Sickinger

Abstract

The human adrenomedullin (ADM) is a 52 amino acid peptide hormone belonging to the calcitonin family of peptides, which plays a major role in the development and regulation of cardiovascular and lymphatic systems. For potential use in clinical applications, we aimed to investigate the fate of the peptide ligand after binding and activation of the adrenomedullin receptor (AM1), a heterodimer consisting of the calcitonin receptor-like receptor (CLR), a G protein-coupled receptor, associated with the receptor activity-modifying protein 2 (RAMP2). Full length and N-terminally shortened ADM peptides were synthesized using Fmoc/tBu solid phase peptide synthesis and site-specifically labeled with the fluorophore carboxytetramethylrhodamine (Tam) either by amide bond formation or copper(I)-catalyzed azide alkyne cycloaddition. For the first time, Tam-labeled ligands allowed the observation of co-internalization of the whole ligand-receptor complex in living cells co-transfected with fluorescent fusion proteins of CLR and RAMP2. Application of a fluorescent probe to track lysosomal compartments revealed that ADM together with the CLR/RAMP2-complex is routed to the degradative pathway. Moreover, we found that the N-terminus of ADM is not...Continue Reading

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Citations

May 12, 2016·Journal of Medicinal Chemistry·Ria SchönauerAnnette G Beck-Sickinger
Feb 6, 2017·Journal of Peptide Science : an Official Publication of the European Peptide Society·Ria SchönauerAnnette G Beck-Sickinger
Mar 10, 2018·Frontiers in Immunology·Christopher GevenPeter Pickkers
Jan 27, 2019·Journal of Peptide Science : an Official Publication of the European Peptide Society·Jan-Patrick FischerAnnette G Beck-Sickinger
Dec 29, 2018·European Journal of Heart Failure·Adriaan A VoorsJaved Butler
Apr 17, 2020·ACS Pharmacology & Translational Science·Michael L GareljaDebbie L Hay

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