State Key Laboratory of Genetic Engineering, Shanghai Key Laboratory of Metabolic Remodeling and Health, Institute of Metabolism and Integrative Biology, Zhongshan Hospital, Fudan University, Shanghai Qi Zhi Institute, Shanghai 200438, China.
State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
J Cell Sci. 2023 Aug 1;136(15). doi: 10.1242/jcs.261319. Epub 2023 Aug 9.
Protein palmitoylation is a post-translational lipid modification of proteins. Accumulating evidence reveals that palmitoylation functions as a sorting signal to direct proteins to destinations; however, the sorting mechanism remains largely unknown. Here, we show that ARF6 plays a general role in targeting palmitoylated proteins from the Golgi to the plasma membrane (PM). Through shRNA screening, we identified ARF6 as the key small GTPase in targeting CD36, a palmitoylated protein, from the Golgi to the PM. We found that the N-terminal myristoylation of ARF6 is required for its binding with palmitoylated CD36, and the GTP-bound form of ARF6 facilitates the delivery of CD36 to the PM. Analysis of stable isotope labeling by amino acids in cell culture revealed that ARF6 might facilitate the sorting of 359 of the 531 palmitoylated PM proteins, indicating a general role of ARF6. Our study has thus identified a sorting mechanism for targeting palmitoylated proteins from the Golgi to the PM.
蛋白质棕榈酰化是蛋白质的一种翻译后脂质修饰。越来越多的证据表明，棕榈酰化作为一种分选信号，指导蛋白质到达特定的目的地；然而，分选机制在很大程度上仍然未知。在这里，我们表明 ARF6 普遍参与将棕榈酰化蛋白从高尔基体靶向到质膜 (PM)。通过 shRNA 筛选，我们确定 ARF6 是将棕榈酰化蛋白 CD36 从高尔基体靶向到 PM 的关键小 GTP 酶。我们发现 ARF6 的 N 端肉豆蔻酰化对于其与棕榈酰化 CD36 的结合是必需的，并且 ARF6 的 GTP 结合形式促进了 CD36 向 PM 的递送。细胞培养中稳定同位素标记的氨基酸分析表明，ARF6 可能有助于分选 531 个棕榈酰化 PM 蛋白中的 359 个，表明 ARF6 具有普遍作用。因此，我们的研究确定了一种从高尔基体到质膜靶向棕榈酰化蛋白的分选机制。
