Ramanoudjame Sridévi M, Esteoulle Lucie, Riché Stéphanie, Margathe Jean-François, Durroux Thierry, Karpenko Iuliia A, Bonnet Dominique
Laboratoire d'Innovation Thérapeutique, Faculté de Pharmacie, UMR 7200 CNRS/Université de Strasbourg, LabEx MEDALIS, Illkirch, France.
CNRS UMR 5203, Institut de Génomique Fonctionnelle and INSERM U1191 and Université de Montpellier, Montpellier, France.
Methods Mol Biol. 2019;1947:137-147. doi: 10.1007/978-1-4939-9121-1_7.
Fluorescence techniques represent a powerful tool to investigate dynamic and functional architecture of GPCRs. Thus, fluorescent GPCR ligands have found various applications in cellular imaging, in the development of binding assays as replacements for radioligands in the study of ligand-receptor but also in receptor-receptor interactions at the cell surface or in native tissues. To extend the applicability of these techniques, the design and the synthesis of fluorescent probes are critical steps. As there are numerous peptide receptors in the GPCR family, fluorescent peptide-based probes are of importance. Herein, we present a convenient method to facilitate the solution-phase fluorescent labeling of peptides which is based on the chemoselective acylation of α-hydrazinopeptides. This approach combines the advantages to use commercially available amine-reactive dyes and very mild conditions that are fully compatible with the chemical sensitivity of the dyes. It gives a rapid access to fluorescent peptidic probes compatible with the time-resolved fluorescence resonance energy transfer (TR-FRET) techniques.
荧光技术是研究G蛋白偶联受体（GPCRs）动态和功能结构的有力工具。因此，荧光GPCR配体已在细胞成像、结合测定的开发中得到了各种应用，可作为放射性配体的替代品用于配体-受体研究，也可用于细胞表面或天然组织中的受体-受体相互作用研究。为了扩展这些技术的适用性，荧光探针的设计和合成是关键步骤。由于GPCR家族中有众多肽受体，基于肽的荧光探针具有重要意义。在此，我们提出一种简便方法，可促进基于α-肼基肽的化学选择性酰化的肽的溶液相荧光标记。该方法结合了使用市售胺反应性染料的优点以及与染料化学敏感性完全兼容的非常温和的条件。它能快速获得与时间分辨荧光共振能量转移（TR-FRET）技术兼容的荧光肽探针。
