Protein lipidation, which regulates numerous biological pathways and plays crucial roles in the pharmaceutical industry, is not encoded by the genetic code but synthesized post-translationally. In the present study, we report a computational approach for designing lipidation mimics that fully recapitulate the biochemical properties of natural lipidation in membrane association and albumin binding. Furthermore, we establish an engineered system for co-translational incorporation of these lipidation mimics into virtually any desired position of proteins in Escherichia coli and mammalian cells. We demonstrate the utility of these length-tunable lipidation mimics in diverse applications, including improving the half-life and activity of therapeutic proteins in living mice, anchoring functional proteins to membrane by substituting natural lipidation, functionally characterizing proteins carrying different lengths of lipidation and determining the plasma membrane-binding capacity of a given compound. Our strategy enables gain-of-function studies of lipidation in hundreds of proteins and facilitates the creation of superior therapeutic candidates.

蛋白质脂化调节许多生物途径并在制药工业中发挥着至关重要的作用，它不是由遗传密码编码，而是在翻译后合成。在本研究中，我们报告了一种设计脂化模拟物的计算方法，该方法完全概括了膜缔合和白蛋白结合中天然脂化的生化特性。此外，我们建立了一个工程系统，用于将这些脂化模拟物共翻译并入大肠杆菌和哺乳动物细胞中蛋白质的几乎任何所需位置。我们展示了这些长度可调的脂化模拟物在不同应用中的效用，包括改善活体小鼠中治疗性蛋白质的半衰期和活性、通过替代天然脂化将功能蛋白锚定到膜上、对携带不同脂化长度的蛋白质进行功能表征以及确定给定化合物的质膜结合能力。我们的策略能够对数百种蛋白质的脂化进行功能获得性研究，并促进优质候选治疗药物的创建。