德国哥廷根大学Silvio O. Rizzoli等研究人员合作发现，一步法纳米级扩展显微术揭示单个蛋白质的形态。这一研究成果于2024年10月9日在线发表在国际学术期刊《自然—生物技术》上。

研究人员将样本扩展至少十倍，用常规荧光染料标记，并使用常规光学显微镜成像，通过分析荧光波动获取视频。一步法纳米级扩展（ONE）显微术能够可视化单个膜蛋白和可溶性蛋白质的形状，分辨率可达约1纳米。

研究人员展示了构象变化的可观测性，例如约17-kDa的钙调蛋白在Ca²⁺结合后发生的构象变化。ONE技术还应用于临床样本，分析帕金森病患者脑脊液中的蛋白质聚集物形态，可能有助于疾病诊断。

这项技术弥合了高分辨率结构生物学技术与光学显微技术之间的差距，为生物学和医学领域的发现提供了新的途径。

据介绍，荧光显微技术的分辨率已经达到亚纳米范围，但该技术仍难以成像单个蛋白质或小分子复合物的形态。

附：英文原文

Title: One-step nanoscale expansion microscopy reveals individual protein shapes

Author: Shaib, Ali H., Chouaib, Abed Alrahman, Chowdhury, Rajdeep, Altendorf, Jonas, Mihaylov, Daniel, Zhang, Chi, Krah, Donatus, Imani, Vanessa, Spencer, Russell K. W., Georgiev, Svilen Veselinov, Mougios, Nikolaos, Monga, Mehar, Reshetniak, Sofiia, Mimoso, Tiago, Chen, Han, Fatehbasharzad, Parisa, Crzan, Dagmar, Saal, Kim-Ann, Alawieh, Mohamad Mahdi, Alawar, Nadia, Eilts, Janna, Kang, Jinyoung, Soleimani, Alireza, Mller, Marcus, Pape, Constantin, Alvarez, Luis, Trenkwalder, Claudia, Mollenhauer, Brit, Outeiro, Tiago F., Kster, Sarah, Preobraschenski, Julia, Becherer, Ute, Moser, Tobias, Boyden, Edward S., Aricescu, A. Radu, Sauer, Markus, Opazo, Felipe, Rizzoli, Silvio O.

Issue&Volume: 2024-10-09

Abstract: The attainable resolution of fluorescence microscopy has reached the subnanometer range, but this technique still fails to image the morphology of single proteins or small molecular complexes. Here, we expand the specimens at least tenfold, label them with conventional fluorophores and image them with conventional light microscopes, acquiring videos in which we analyze fluorescence fluctuations. One-step nanoscale expansion (ONE) microscopy enables the visualization of the shapes of individual membrane and soluble proteins, achieving around 1-nm resolution. We show that conformational changes are readily observable, such as those undergone by the ~17-kDa protein calmodulin upon Ca2+ binding. ONE is also applied to clinical samples, analyzing the morphology of protein aggregates in cerebrospinal fluid from persons with Parkinson disease, potentially aiding disease diagnosis. This technology bridges the gap between high-resolution structural biology techniques and light microscopy, providing new avenues for discoveries in biology and medicine.

DOI: 10.1038/s41587-024-02431-9

Source: https://www.nature.com/articles/s41587-024-02431-9