Protein lipidation dramatically affects protein structure, localization, and trafficking via remodeling protein–membrane and protein–protein interactions through hydrophobic lipid moieties. Understanding the biosynthesis of lipidated proteins, whether natural ones or mimetics, is crucial for reconstructing, validating, and studying the molecular mechanisms and biological functions of protein lipidation. In this Perspective, we first provide an overview of the natural enzymatic biosynthetic pathways of protein lipidation in mammalian cells, focusing on the enzymatic machineries and their chemical linkages. We then discuss strategies to biosynthesize protein lipidation in mammalian cells by engineering modification machineries and substrates. Additionally, we explore site-specific protein lipidation biosynthesis in vitro via enzyme-mediated ligations and in vivo primarily through genetic code expansion strategies. We also discuss the use of small molecule tools to modulate the process of protein lipidation biosynthesis. Finally, we provide concluding remarks and discuss future directions for the biosynthesis and applications of protein lipidation.

中文翻译：

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蛋白质脂质化的生物合成和应用


蛋白质脂质化通过疏水性脂质部分重塑蛋白质-膜和蛋白质-蛋白质相互作用，从而显著影响蛋白质结构、定位和运输。了解脂质化蛋白质的生物合成，无论是天然蛋白质还是模拟物，对于重建、验证和研究蛋白质脂质化的分子机制和生物学功能至关重要。在这个 Perspective 中，我们首先概述了哺乳动物细胞中蛋白质脂质化的天然酶生物合成途径，重点介绍了酶机制及其化学键。然后，我们讨论了通过工程修饰机制和底物在哺乳动物细胞中生物合成蛋白质脂质化的策略。此外，我们通过酶介导的连接在体外探索位点特异性蛋白质脂质化生物合成，主要通过遗传密码扩增策略在体内探索。我们还讨论了使用小分子工具来调节蛋白质脂质化生物合成过程。最后，我们提供了结论，并讨论了蛋白质脂质化生物合成和应用的未来方向。