Kol Matthijs, Novak Alexander J E, Morstein Johannes, Schröer Christian, Sokoya Tolulope, Mensing Svenja, Korneev Sergei M, Trauner Dirk, Holthuis Joost C M
Molecular Cell Biology Division, Department of Biology/Chemistry, Osnabrück University, 49076 Osnabrück, Germany.
Center for Cellular Nanoanalytics, Osnabrück University, Artilleriestraße 77, 49076 Osnabrück, Germany.
bioRxiv. 2024 Oct 24:2024.10.24.619506. doi: 10.1101/2024.10.24.619506.
Sphingolipid metabolism comprises a complex interconnected web of enzymes, metabolites and modes of regulation that influence a wide range of cellular and physiological processes. Deciphering the biological relevance of this network is challenging as numerous intermediates of sphingolipid metabolism are short-lived molecules with often opposing biological activities. Here, we introduce clickable, azobenzene-containing sphingosines, termed s, as light-sensitive substrates for sphingolipid biosynthesis. Photo-isomerization of the azobenzene moiety enables reversible switching between a straight - and curved -form of the lipid's hydrocarbon tail. Combining enzyme assays with metabolic labeling studies, we demonstrate that -to- isomerization of s profoundly stimulates their metabolic conversion by ceramide synthases and downstream sphingomyelin synthases. These light-induced changes in sphingolipid production rates are acute, reversible, and can be implemented with great efficiency in living cells. Our findings establish s as versatile tools with unprecedented opportunities to manipulate sphingolipid biosynthesis and function with the spatiotemporal precision of light.
鞘脂代谢由酶、代谢物和调节模式构成一个复杂的相互关联网络，影响着广泛的细胞和生理过程。解读该网络的生物学意义具有挑战性，因为鞘脂代谢的众多中间产物都是寿命短暂的分子，且往往具有相反的生物学活性。在此，我们引入了可点击的含偶氮苯的鞘氨醇（称为s），作为鞘脂生物合成的光敏底物。偶氮苯部分的光异构化可使脂质烃尾在直链和弯曲形式之间进行可逆切换。结合酶分析和代谢标记研究，我们证明s从反式到顺式的异构化极大地刺激了它们被神经酰胺合酶和下游鞘磷脂合酶的代谢转化。鞘脂生成速率的这些光诱导变化是快速、可逆的，并且能够在活细胞中高效实现。我们的研究结果将s确立为多功能工具，为以前所未有的时空精度操纵鞘脂生物合成和功能提供了机会。
