小柯机器人

瑞典皇家理工学院Ilaria Testa团队近期取得重要工作进展，他们研究利用可逆转换的蛋白将荧光各向异性扩展到大型复合体上。相关研究工作2022年10月10日在线发表于《自然—生物技术》杂志上。

研究人员描述了具有可逆开关切换状态的选择性时间分辨各向异性（STARSS），将可观察的质量范围扩大了三个数量级以上。STARSS是基于可转换荧光蛋白的长效可逆分子转换，以解决大型复合体相对缓慢的旋转扩散性。研究人员以STARSS探测了细胞中几个分子复合体的旋转流动性，包括染色质、逆转录病毒Gag晶格和活性调节的细胞骨架相关蛋白寡聚体。由于STARSS可以探测任意大的结构，它普遍适用于整个人类蛋白质组。

据介绍，大分子复合物的形成可以通过使用时间分辨的荧光各向异性检测旋转流动性的变化来测量。然而，这种方法仅限于相对较小的分子（~0.1-30 kDa），排除了大部分人类蛋白质组及其复合物。

附：英文原文

Title: Extending fluorescence anisotropy to large complexes using reversibly switchable proteins

Author: Volpato, Andrea, Ollech, Dirk, Alvelid, Jonatan, Damenti, Martina, Mller, Barbara, York, Andrew G, Ingaramo, Maria, Testa, Ilaria

Issue&Volume: 2022-10-10

Abstract: The formation of macromolecular complexes can be measured by detection of changes in rotational mobility using time-resolved fluorescence anisotropy. However, this method is limited to relatively small molecules (~0.1–30kDa), excluding the majority of the human proteome and its complexes. We describe selective time-resolved anisotropy with reversibly switchable states (STARSS), which overcomes this limitation and extends the observable mass range by more than three orders of magnitude. STARSS is based on long-lived reversible molecular transitions of switchable fluorescent proteins to resolve the relatively slow rotational diffusivity of large complexes. We used STARSS to probe the rotational mobility of several molecular complexes in cells, including chromatin, the retroviral Gag lattice and activity-regulated cytoskeleton-associated protein oligomers. Because STARSS can probe arbitrarily large structures, it is generally applicable to the entire human proteome.

DOI: 10.1038/s41587-022-01489-7

Source: https://www.nature.com/articles/s41587-022-01489-7

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex