美国华盛顿大学David Baker等研究人员合作开发出，以亚细胞分辨率检测内源性Ras活性和信号作用因子的计算设计传感器。相关论文于2024年1月25日在线发表于国际学术期刊《自然—生物技术》。

研究人员利用计算蛋白质设计生成了细胞内Ras活性传感器（基于LOCKR的Ras活性传感器，Ras-LOCKR-S），和Ras信号环境的近距离标记物（基于LOCKR的，且依赖Ras活性的近距离标记物，Ras-LOCKR-PL）。这些工具可以检测内源性Ras活性，并以亚细胞分辨率对周围环境进行标记。

通过在人类癌细胞系中使用这些传感器，研究人员确定了致癌EML4-Alk颗粒中的Ras相互作用蛋白，并发现SAM68（Src-Associated in Mitosis 68-kDa）蛋白特异性地增强了颗粒中的Ras活性。对内源性Ras活性进行亚细胞定位的能力将加深人们对健康和疾病中Ras功能的了解，并可能提出潜在的治疗策略。

据了解，由于缺乏将传感器灵敏度与目标物生理浓度范围相匹配的策略，基因编码生物传感器在感知信号蛋白活性方面的应用受到了阻碍。

附：英文原文

Title: Computationally designed sensors detect endogenous Ras activity and signaling effectors at subcellular resolution

Author: Zhang, Jason Z., Nguyen, William H., Greenwood, Nathan, Rose, John C., Ong, Shao-En, Maly, Dustin J., Baker, David

Issue&Volume: 2024-01-25

Abstract: The utility of genetically encoded biosensors for sensing the activity of signaling proteins has been hampered by a lack of strategies for matching sensor sensitivity to the physiological concentration range of the target. Here we used computational protein design to generate intracellular sensors of Ras activity (LOCKR-based Sensor for Ras activity (Ras-LOCKR-S)) and proximity labelers of the Ras signaling environment (LOCKR-based, Ras activity-dependent Proximity Labeler (Ras-LOCKR-PL)). These tools allow the detection of endogenous Ras activity and labeling of the surrounding environment at subcellular resolution. Using these sensors in human cancer cell lines, we identified Ras-interacting proteins in oncogenic EML4-Alk granules and found that Src-Associated in Mitosis 68-kDa (SAM68) protein specifically enhances Ras activity in the granules. The ability to subcellularly localize endogenous Ras activity should deepen our understanding of Ras function in health and disease and may suggest potential therapeutic strategies.

DOI: 10.1038/s41587-023-02107-w

Source: https://www.nature.com/articles/s41587-023-02107-w