PDF(Pubmed)
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.
We describe Trp‐BODIPY PLUS as a new acid‐resistant building block for straightforward solid‐phase synthesis of fluorogenic peptides. We demonstrate the utility of Trp‐BODIPY PLUS with the first fluorogenic probe for direct visualization and quantitative analysis of the expression and localization of the G protein‐coupled receptors 54 (GPR54 or kisspeptin) in human cells and intact mouse pancreatic islets.
We describe Trp‐BODIPY PLUS as a new acid‐resistant building block for straightforward solid‐phase synthesis of fluorogenic peptides. We demonstrate the utility of Trp‐BODIPY PLUS with the first fluorogenic probe for direct visualization and quantitative analysis of the expression and localization of the G protein‐coupled receptors 54 (GPR54 or kisspeptin) in human cells and intact mouse pancreatic islets.
摘要：
G蛋白偶联kisspeptin受体（GPR54或KISS1R）是生殖中的重要介质，代谢和癌症生物学;然而，有有限的荧光探针或抗体直接成像的这些受体在细胞和完整的组织，这可以帮助询问他们的多种生物学角色。在这里，我们描述了一种新的耐酸的基于BODIPY的氨基酸（Trp-BODIPYPLUS）的合理设计和表征，及其用于固相合成荧光生物活性肽的实现。Trp-BODIPYPLUS保留了短线性和环状肽的结合能力，并在靶标结合时显示出明显的开启荧光发射，用于无洗涤成像。最后,我们使用Trp-BODIPYPLUS制备了一些首批基于kisspeptin的荧光探针,并通过荧光成像观察了GPR54受体在人细胞和整个小鼠胰岛中的表达和定位.
我们将Trp-BODIPYPLUS描述为一种新的耐酸结构单元，用于直接固相合成荧光肽。我们证明了Trp-BODIPYPLUS与第一个荧光探针的实用性，可直接可视化和定量分析人细胞和完整小鼠胰岛中G蛋白偶联受体54（GPR54或kisspeptin）的表达和定位。
我们将Trp-BODIPYPLUS描述为一种新的耐酸结构单元，用于直接固相合成荧光肽。我们证明了Trp-BODIPYPLUS与第一个荧光探针的实用性，可直接可视化和定量分析人细胞和完整小鼠胰岛中G蛋白偶联受体54（GPR54或kisspeptin）的表达和定位。
