MINFLUX dissects the unimpeded walking of kinesin-1

• Verfasst von: Wolff, Jan Otto [VerfasserIn]
• Verfasst von: Scheiderer, Jan Lukas [VerfasserIn]
• Verfasst von: Engelhardt, Tobias [VerfasserIn]
• Verfasst von: Engelhardt, Johann [VerfasserIn]
• Verfasst von: Matthias, Jessica [VerfasserIn]
• Verfasst von: Hell, Stefan [VerfasserIn]
• Titel: MINFLUX dissects the unimpeded walking of kinesin-1
• Verf.angabe: Jan O. Wolff, Lukas Scheiderer, Tobias Engelhardt, Johann Engelhardt, Jessica Matthias, Stefan W. Hell
• E-Jahr: 2023
• Jahr: March 10, 2023
• Umfang: 7 S.
• Fussnoten: Gesehen am 13.03.2023
• Titel Quelle: Enthalten in: Science
• Ort Quelle: Washington, DC : American Association for the Advancement of Science, 1880
• Jahr Quelle: 2023
• Band/Heft Quelle: 379(2023), 6636, Seite 1004-1010
• ISSN Quelle: 1095-9203
• DOI: doi:10.1126/science.ade2650
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1839010185

Abstract

We introduce an interferometric MINFLUX microscope that records protein movements with up to 1.7 nanometer per millisecond spatiotemporal precision. Such precision has previously required attaching disproportionately large beads to the protein, but MINFLUX requires the detection of only about 20 photons from an approximately 1-nanometer-sized fluorophore. Therefore, we were able to study the stepping of the motor protein kinesin-1 on microtubules at up to physiological adenosine-5′-triphosphate (ATP) concentrations. We uncovered rotations of the stalk and the heads of load-free kinesin during stepping and showed that ATP is taken up with a single head bound to the microtubule and that ATP hydrolysis occurs when both heads are bound. Our results show that MINFLUX quantifies (sub)millisecond conformational changes of proteins with minimal disturbance.