Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, California 92093, United States.
Department of Bioengineering, University of California San Diego, La Jolla, California 92093, United States.
J Am Chem Soc. 2020 Oct 21;142(42):17887-17891. doi: 10.1021/jacs.0c06652. Epub 2020 Oct 12.
The single-chained sphingolipid sphingosine is an essential structural lipid and signaling molecule. Abnormal sphingosine metabolism is observed in several diseases, including cancer, diabetes, and Alzheimer's. Despite its biological importance, there is a lack of tools for detecting sphingosine in living cells. This is likely due to the broader challenge of developing highly selective and live-cell compatible affinity probes for hydrophobic lipid species. In this work, we have developed a small molecule fluorescent turn-on probe for labeling sphingosine in living cells. We demonstrate that this probe exhibits a dose-dependent response to sphingosine and is able to detect endogenous pools of sphingosine. Using our probe, we successfully detected sphingosine accumulation in cells from patients with Niemann-Pick type C1 (NPC1), a lipid transport disorder in which increased sphingosine mediates disease progression. This work provides a simple and accessible method for the detection of sphingosine and should facilitate study of this critical signaling lipid in biology and disease.
单链神经鞘脂神经醇是一种必需的结构脂质和信号分子。几种疾病（包括癌症、糖尿病和阿尔茨海默病）中都观察到神经醇代谢异常。尽管它具有重要的生物学意义，但缺乏用于检测活细胞中神经醇的工具。这可能是由于开发用于疏水性脂质种类的高选择性和活细胞兼容亲和探针的更广泛挑战所致。在这项工作中，我们开发了一种小分子荧光开启探针，用于标记活细胞中的神经醇。我们证明该探针对神经醇表现出剂量依赖性响应，并且能够检测内源性神经醇池。使用我们的探针，我们成功地检测到 NPC1 患者细胞中神经醇的积累，NPC1 是一种脂质转运障碍，其中增加的神经醇介导疾病进展。这项工作为神经醇的检测提供了一种简单易用的方法，应有助于在生物学和疾病中研究这种关键的信号脂质。
