2024-10-10 マックス・プランク研究所
Polyprolines of different lengths, relatively stiff polypeptides, served as intramolecular “nanometer rulers” to demonstrate the highest MINFLUX resolutions in the Förster resonance energy transfer (FRET) distance range. The 2-sigma ellipses show the measurement uncertainty of the individual positions.
© MPI f. Multidisciplinary Sciences/ Steffen J. Sahl
＜関連情報＞
- https://www.mpg.de/23554462/1010-bich-direct-optical-measuerement-of-intramolecular-distances-with-angstrom-precision-17216463-x
- https://www.science.org/doi/10.1126/science.adj7368
オングストローム精度で分子内距離を光学的に直接測定 Direct optical measurement of intramolecular distances with angstrom precision
Steffen J. Sahl, Jessica Matthias, Kaushik Inamdar, Michael Weber, […], and Stefan W. Hell
Science Published:10 Oct 2024
DOI:https://doi.org/10.1126/science.adj7368
Editor’s summary
Measuring distances directly at the nanometer scale is a challenge for optical techniques, even for those using subdiffraction-resolution fluorescence microscopy. Sahl et al. refined an optical approach called MINFLUX such that they could measure precise intramolecular distances in the 1- to 10-nanometer range and below 1 nanometer for molecules with a tilt. Using a polyproline ruler, the authors demonstrate resolution of fluorophores with known single-digit nanometer spacing. They applied this approach to inter- and intramacromolecular measurements of proteins labeled with photoactivatable dyes, including distances too short for current indirect methods. Imaging experiments demonstrated the potential of this technique to study protein-protein interactions in cells. —Michael A. Funk
Abstract
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.
