Agam, Ganesh and Gebhardt, Christian and Popara, Milana and Maechtel, Rebecca and Folz, Julian and Ambrose, Benjamin and Chamachi, Neharika and Chung, Sang Yoon and Craggs, Timothy D. and de Boer, Marijn and Grohmann, Dina and Ha, Taekjip and Hartmann, Andreas and Hendrix, Jelle and Hirschfeld, Verena and Huebner, Christian G. and Hugel, Thorsten and Kammerer, Dominik and Kang, Hyun-Seo and Kapanidis, Achillefs N. and Krainer, Georg and Kramm, Kevin and Lemke, Edward A. and Lerner, Eitan and Margeat, Emmanuel and Martens, Kirsten and Michaelis, Jens and Mitra, Jaba and Munoz, Gabriel G. Moya and Quast, Robert B. and Robb, Nicole C. and Sattler, Michael and Schlierf, Michael and Schneider, Jonathan and Schroeder, Tim and Sefer, Anna and Tan, Piau Siong and Thurn, Johann and Tinnefeld, Philip and van Noort, John and Weiss, Shimon and Wendler, Nicolas and Zijlstra, Niels and Barth, Anders and Seidel, Claus A. M. and Lamb, Don C. and Cordes, Thorben (2023) Reliability and accuracy of single-molecule FRET studies for characterization of structural dynamics and distances in proteins. NATURE METHODS, 20 (4). ISSN 1548-7091, 1548-7105
Full text not available from this repository. (Request a copy)Abstract
Single-molecule Forster-resonance energy transfer (smFRET) experiments allow the study of biomolecular structure and dynamics in vitro and in vivo. We performed an international blind study involving 19 laboratories to assess the uncertainty of FRET experiments for proteins with respect to the measured FRET efficiency histograms, determination of distances, and the detection and quantification of structural dynamics. Using two protein systems with distinct conformational changes and dynamics, we obtained an uncertainty of the FRET efficiency <= 0.06, corresponding to an interdye distance precision of <= 2 angstrom and accuracy of <= 5 angstrom. We further discuss the limits for detecting fluctuations in this distance range and how to identify dye perturbations. Our work demonstrates the ability of smFRET experiments to simultaneously measure distances and avoid the averaging of conformational dynamics for realistic protein systems, highlighting its importance in the expanding toolbox of integrative structural biology. An international blind study confirms that smFRET measurements on dynamic proteins are highly reproducible across instruments, analysis procedures and timescales, further highlighting the promise of smFRET for dynamic structural biology.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RESONANCE ENERGY-TRANSFER; ALTERNATING-LASER EXCITATION; NANO-POSITIONING SYSTEM; CONFORMATIONAL DYNAMICS; ORIENTATIONAL FREEDOM; LIFETIME DISTRIBUTION; PHOTON DISTRIBUTION; LIGAND-BINDING; RNA; SPECTROSCOPY; |
| Subjects: | 500 Science > 550 Earth sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Dina Grohmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 30 Jan 2024 07:34 |
| Last Modified: | 30 Jan 2024 07:34 |
| URI: | https://pred.uni-regensburg.de/id/eprint/60958 |
Actions (login required)
 |
View Item |
