Max Planck Institute for the Science of Light, Erlangen, Germany.
Department of Physics, Friedrich Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany.
Nat Methods. 2017 Feb;14(2):141-144. doi: 10.1038/nmeth.4141. Epub 2017 Jan 9.
We introduce Cryogenic Optical Localization in 3D (COLD), a method to localize multiple fluorescent sites within a single small protein with Angstrom resolution. We demonstrate COLD by determining the conformational state of the cytosolic Per-ARNT-Sim domain from the histidine kinase CitA of Geobacillus thermodenitrificans and resolving the four biotin sites of streptavidin. COLD provides quantitative 3D information about small- to medium-sized biomolecules on the Angstrom scale and complements other techniques in structural biology.
我们介绍了 Cryogenic Optical Localization in 3D (COLD)，这是一种在单个小蛋白内定位多个荧光点的方法，分辨率可达埃。我们通过确定来自 Geobacillus thermodenitrificans 的组氨酸激酶 CitA 的胞质 Per-ARNT-Sim 结构域的构象状态，并解析链霉亲和素的四个生物素结合位点，证明了 COLD 的可行性。COLD 可提供关于埃尺度中小至中等大小生物分子的定量 3D 信息，并补充结构生物学中的其他技术。
