Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen 37077, Germany.
Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg 69120, Germany.
Science. 2024 Oct 11;386(6718):180-187. doi: 10.1126/science.adj7368. Epub 2024 Oct 10.
Optical investigations of nanometer distances between proteins, their subunits, or other biomolecules have been the exclusive prerogative of Förster resonance energy transfer (FRET) microscopy for decades. In this work, we show that MINFLUX fluorescence nanoscopy measures intramolecular distances down to 1 nanometer-and in planar projections down to 1 angstrom-directly, linearly, and with angstrom precision. Our method was validated by quantifying well-characterized 1- to 10-nanometer distances in polypeptides and proteins. Moreover, we visualized the orientations of immunoglobulin subunits, applied the method in human cells, and revealed specific configurations of a histidine kinase PAS domain dimer. Our results open the door for examining proximities and interactions by direct position measurements at the intramacromolecular scale.
几十年来，对蛋白质、其亚基或其他生物分子之间纳米距离的光学研究一直是福斯特共振能量转移（FRET）显微镜的专属特权。在这项工作中，我们表明 MINFLUX 荧光纳米显微镜可以直接、线性和以埃精度测量低至 1 纳米的分子内距离，并且在平面投影中低至 1 埃。我们的方法通过定量多肽和蛋白质中特征良好的 1 到 10 纳米距离得到了验证。此外，我们还可视化了免疫球蛋白亚基的取向，将该方法应用于人类细胞，并揭示了组氨酸激酶 PAS 结构域二聚体的特定构型。我们的结果为通过在分子内尺度上进行直接位置测量来检查接近程度和相互作用开辟了道路。
