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Acid-Resistant BODIPY Amino Acids for Peptide-Based Fluorescence Imaging of GPR54 Receptors in Pancreatic Islets
Lorena Mendive-Tapia1, Laia Miret-Casals2, Nicole D Barth1
1Centre for Inflammation Research, The University of Edinburgh, EH16 4TJ, Edinburgh, UK.
Abstract:
The G protein-coupled kisspeptin receptor (GPR54 or KISS1R) is an important mediator in reproduction, metabolism and cancer biology; however, there are limited fluorescent probes or antibodies for direct imaging of these receptors in cells and intact tissues, which can help to interrogate their multiple biological roles. Herein, we describe the rational design and characterization of a new acid-resistant BODIPY-based amino acid (Trp-BODIPY PLUS), and its implementation for solid-phase synthesis of fluorescent bioactive peptides. Trp-BODIPY PLUS retains the binding capabilities of both short linear and cyclic peptides and displays notable turn-on fluorescence emission upon target binding for wash-free imaging. Finally, we employed Trp-BODIPY PLUS to prepare some of the first fluorogenic kisspeptin-based probes and visualized the expression and localization of GPR54 receptors in human cells and in whole mouse pancreatic islets by fluorescence imaging.
Insights
Researchers developed a new fluorescent probe for imaging the kisspeptin receptor (GPR54). This tool enables visualization of GPR54 in cells and tissues, advancing research in reproduction, metabolism, and cancer.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- The kisspeptin receptor (GPR54 or KISS1R) is crucial for reproduction, metabolism, and cancer.
- Limited availability of fluorescent probes and antibodies hinders direct imaging of GPR54.
- Understanding GPR54's roles requires advanced imaging tools for cellular and tissue analysis.
Purpose of the Study:
- To design and characterize a novel acid-resistant fluorescent amino acid for peptide synthesis.
- To develop fluorogenic kisspeptin-based probes for GPR54 imaging.
- To visualize GPR54 expression and localization in human cells and mouse pancreatic islets.
Main Methods:
- Rational design and synthesis of an acid-resistant BODIPY-based amino acid (Trp-BODIPY PLUS).
- Solid-phase synthesis of fluorescent bioactive peptides using Trp-BODIPY PLUS.
- Preparation of fluorogenic kisspeptin analogs and fluorescence imaging experiments.
Main Results:
- Trp-BODIPY PLUS demonstrated retained peptide binding capabilities and turn-on fluorescence upon target binding.
- Wash-free imaging was achieved due to the probe's properties.
- The first fluorogenic kisspeptin probes were successfully synthesized and used to visualize GPR54 in human cells and mouse pancreatic islets.
Conclusions:
- Trp-BODIPY PLUS is a versatile tool for creating fluorescent bioactive peptides.
- The developed kisspeptin probes enable effective visualization of GPR54 receptors.
- This work provides novel imaging capabilities for studying GPR54 in various biological contexts.
