de Moliner Fabio, Konieczna Zuzanna, Mendive-Tapia Lorena, Saleeb Rebecca S, Morris Katie, Gonzalez-Vera Juan Antonio, Kaizuka Takeshi, Grant Seth G N, Horrocks Mathew H, Vendrell Marc
Centre for Inflammation Research The University of Edinburgh UK.
EaStCHEM School of Chemistry The University of Edinburgh UK.
Angew Chem Weinheim Bergstr Ger. 2023 Jan 23;135(4):e202216231. doi: 10.1002/ange.202216231. Epub 2022 Dec 14.
The multiple applications of super-resolution microscopy have prompted the need for minimally invasive labeling strategies for peptide-guided fluorescence imaging. Many fluorescent reporters display limitations (e.g., large and charged scaffolds, non-specific binding) as building blocks for the construction of fluorogenic peptides. Herein we have built a library of benzodiazole amino acids and systematically examined them as reporters for background-free fluorescence microscopy. We have identified amine-derivatized benzoselenadiazoles as scalable and photostable amino acids for the straightforward solid-phase synthesis of fluorescent peptides. Benzodiazole amino acids retain the binding capabilities of bioactive peptides and display excellent signal-to-background ratios. Furthermore, we have demonstrated their application in peptide-PAINT imaging of postsynaptic density protein-95 nanoclusters in the synaptosomes from mouse brain tissues.
超分辨率显微镜的多种应用促使人们需要用于肽引导荧光成像的微创标记策略。作为构建荧光肽的基础材料，许多荧光报告基团存在局限性（例如，支架大且带电荷、非特异性结合）。在此，我们构建了一个苯并二氮杂环氨基酸库，并系统地将它们作为无背景荧光显微镜的报告基团进行了研究。我们已确定胺衍生化的苯并硒二唑为可扩展且光稳定的氨基酸，可用于直接固相合成荧光肽。苯并二氮杂环氨基酸保留了生物活性肽的结合能力，并显示出优异的信噪比。此外，我们还展示了它们在小鼠脑组织突触体中突触后密度蛋白95纳米簇的肽-PAINT成像中的应用。
