Direct optical measurement of intramolecular distances with angstrom precision

• 成果类型: Article
• 署名作者: Sahl, Steffen J.; Matthias, Jessica; Inamdar, Kaushik; Weber, Michael; Khan, Taukeer A.; Brueser, Christian; Jakobs, Stefan; Becker, Stefan; Griesinger, Christian; Broichhagen, Johannes; Hell, Stefan W.
• 署名单位: Max Planck Society; University of Gottingen; UNIVERSITY GOTTINGEN HOSPITAL; University of Gottingen; Max Planck Society
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-9159
• DOI: 10.1126/science.adj7368
• 发表日期: 2024-10-01
• 页码: 180-187
• 关键词: single-molecule fret; energy-transfer; localization; accuracy; polyproline; colocalization; microscopy; binding; lamins

摘要

Optical investigations of nanometer distances between proteins, their subunits, or other biomolecules have been the exclusive prerogative of F & ouml;rster 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.