Department of Pharmacology, Toxicology and Medicinal Chemistry, Unit of Pharmaceutical Chemistry (Associated Unit to CSIC). Faculty of Pharmacy and Food Sciences. University of Barcelona (UB), Joan XXIII 27-31, 08028 Barcelona, Spain.
Org Biomol Chem. 2021 Mar 21;19(11):2456-2467. doi: 10.1039/d1ob00113b. Epub 2021 Mar 2.
The suitability as FRET probes of two bichromophoric 1-deoxydihydroceramides containing a labelled spisulosine derivative as a sphingoid base and two differently ω-labelled fluorescent palmitic acids has been evaluated. The ceramide synthase (CerS) catalyzed metabolic incorporation of ω-azido palmitic acid into the above labeled spisulosine to render the corresponding ω-azido 1-deoxyceramide has been studied in several cell lines. In addition, the strain-promoted click reaction between this ω-azido 1-deoxyceramide and suitable fluorophores has been optimized to render the target bichromophoric 1-deoxydihydroceramides. These results pave the way for the development of FRET-based assays as a new tool to study sphingolipid metabolism.
已经评估了两种双生色 1-脱氧神经酰胺作为 FRET 探针的适用性，它们含有一个标记的螺旋甾醇衍生物作为神经酰胺碱基和两个不同ω-标记的荧光棕榈酸。已经研究了神经酰胺合酶 (CerS) 在几种细胞系中催化 ω-叠氮棕榈酸的代谢掺入到上述标记的螺旋甾醇中，以生成相应的 ω-叠氮 1-脱氧神经酰胺。此外，优化了这种 ω-叠氮 1-脱氧神经酰胺与合适荧光团之间的应变促进点击反应，以生成目标双生色 1-脱氧二氢神经酰胺。这些结果为开发基于 FRET 的测定方法作为研究鞘脂代谢的新工具铺平了道路。
