美国耶鲁大学Kallol Gupta课题组表明，一种蛋白质组范围的定量平台可用于将膜蛋白提取到原生纳米盘中，并实现纳米尺度空间分辨。相关论文于2024年11月28日在线发表于国际学术期刊《自然—方法学》。

研究人员开发了一种膜蛋白组范围的平台，该平台利用膜活性聚合物库，能够快速、空间分辨地从细胞膜中提取目标膜蛋白，并将其直接嵌入保持局部膜环境的原生纳米盘中。研究人员还提供了一个开放访问的数据库，量化了2065种独特哺乳动物膜蛋白的聚合物特异性提取效率，并为每种膜蛋白提供了最优化的提取条件。

为了验证，研究人员展示了如何利用这一资源从不同的细胞器膜中高效、纯净地提取和纯化目标膜蛋白。此外，通过举例两种突触囊泡膜蛋白，研究人员还展示了如何将该数据库扩展，以捕获过表达的多蛋白复合物。研究人员预计，这些公开资源将帮助跨学科的研究人员高效地捕获含有目标膜蛋白的膜“纳米勺”，并与结构、功能和生物分析方法结合使用。

据悉，天然膜环境深刻影响着膜蛋白生物学的各个方面。尽管如此，最常用的研究膜蛋白的方法是使用去污剂破坏并去除这一重要的膜背景，这阻碍了人们解读局部分子背景及其作用的能力。

附：英文原文

Title: A proteome-wide quantitative platform for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs

Author: Brown, Caroline, Ghosh, Snehasish, McAllister, Rachel, Kumar, Mukesh, Walker, Gerard, Sun, Eric, Aman, Talat, Panda, Aniruddha, Kumar, Shailesh, Li, Wenxue, Coleman, Jeff, Liu, Yansheng, Rothman, James E., Bhattacharyya, Moitrayee, Gupta, Kallol

Issue&Volume: 2024-11-28

Abstract: The native membrane environment profoundly influences every aspect of membrane protein (MP) biology. Despite this, the most prevalent method of studying MPs uses detergents to disrupt and remove this vital membrane context, impeding our ability to decipher the local molecular context and its effect. Here we develop a membrane proteome-wide platform that enables rapid spatially resolved extraction of target MPs directly from cellular membranes into native nanodiscs that maintain the local membrane context, using a library of membrane-active polymers. We accompany this with an open-access database that quantifies the polymer-specific extraction efficiency for 2,065 unique mammalian MPs and provides the most optimized extraction condition for each. To validate, we demonstrate how this resource can enable rapid extraction and purification of target MPs from different organellar membranes with high efficiency and purity. Further, we show how the database can be extended to capture overexpressed multiprotein complexes by taking two synaptic vesicle MPs. We expect these publicly available resources to empower researchers across disciplines to efficiently capture membrane ‘nano-scoops’ containing a target MP and interface with structural, functional and bioanalytical approaches.

DOI: 10.1038/s41592-024-02517-x

Source:https://www.nature.com/articles/s41592-024-02517-x