Reliability and accuracy of single-molecule FRET studies for characterization of structural dynamics and distances in proteins
| العنوان: | Reliability and accuracy of single-molecule FRET studies for characterization of structural dynamics and distances in proteins |
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| المؤلفون: | Agam, Ganesh, Gebhardt, Christian, Popara, Milana, Mächtel, Rebecca, Folz, Julian, Ambrose, Benjamin, Chamachi, Neharika, Chung, Sang Yoon, Craggs, Timothy D, De Boer, Marijn, Grohmann, Dina, Ha, Taekjip, Hartmann, Andreas, Hendrix, Jelle, Hirschfeld, Verena, Hübner, Christian G, Hugel, Thorsten, Kammerer, Dominik, Kang, Hyun-Seo, Kapanidis, Achillefs N, Krainer, Georg, Kramm, Kevin, Lemke, Edward A, Lerner, Eitan, Margeat, Emmanuel, Martens, Kirsten, Michaelis, Jens, Mitra, Jaba, Moya Muñoz, Gabriel G, Quast, Robert B, Robb, Nicole C, Sattler, Michael, Schlierf, Michael, Schneider, Jonathan, Schröder, Tim, Sefer, Anna, Tan, Piau Siong, Thurn, Johann, Tinnefeld, Philip, Van Noort, John, Weiss, Shimon, Wendler, Nicolas, Zijlstra, Niels, Barth, Anders, Seidel, Claus AM, Lamb, Don C, Cordes, Thorben |
| المساهمون: | Agam, Ganesh [0000-0003-1623-4920], Gebhardt, Christian [0000-0002-5834-6579], Popara, Milana [0000-0003-2626-6096], Folz, Julian [0000-0001-8232-8787], Ambrose, Benjamin [0000-0002-5046-2532], Chamachi, Neharika [0000-0002-4657-9092], Craggs, Timothy D [0000-0002-7121-0609], Grohmann, Dina [0000-0002-0570-2517], Ha, Taekjip [0000-0003-2195-6258], Hartmann, Andreas [0000-0002-2213-2763], Hendrix, Jelle [0000-0001-5731-1297], Hugel, Thorsten [0000-0003-3292-4569], Kang, Hyun-Seo [0000-0003-4029-5096], Krainer, Georg [0000-0002-9626-7636], Lemke, Edward A [0000-0002-0634-0503], Lerner, Eitan [0000-0002-3791-5277], Margeat, Emmanuel [0000-0001-6063-6420], Michaelis, Jens [0000-0002-2739-4172], Moya Muñoz, Gabriel G [0000-0003-2797-2005], Quast, Robert B [0000-0001-8166-6952], Schlierf, Michael [0000-0002-6209-2364], Schröder, Tim [0000-0002-2474-5395], Thurn, Johann [0000-0001-6370-7052], van Noort, John [0000-0002-9094-5598], Weiss, Shimon [0000-0002-0720-5426], Wendler, Nicolas [0000-0002-5436-3929], Barth, Anders [0000-0003-3671-3072], Seidel, Claus AM [0000-0002-5171-149X], Lamb, Don C [0000-0002-0232-1903], Cordes, Thorben [0000-0002-8598-5499], Apollo - University of Cambridge Repository, Agam, Ganesh/0000-0003-1623-4920, Popara, Milana/0000-0003-2626-6096, Chamachi, Neharika/0000-0002-4657-9092, Quast, Robert, Benjamin/0000-0001-8166-6952, Agam, Ganesh, Gebhardt, Christian, Popara, Milana, Maechtel, Rebecca, Folz, Julian, Ambrose, Benjamin, Chamachi, Neharika, Chung, Sang Yoon, Craggs, Timothy D., de Boer, Marijn, Grohmann, Dina, Ha, Taekjip, Hartmann, Andreas, HENDRIX, Jelle, Hirschfeld, Verena, Huebner, Christian G., Hugel, Thorsten, Kammerer, Dominik, Kang, Hyun-Seo, Kapanidis, Achillefs N., Krainer, Georg, Kramm, Kevin, Lemke, Edward A., Lerner, Eitan, Margeat, Emmanuel, Martens , Kirsten, Michaelis, Jens, Mitra, Jaba, Munoz, Gabriel G. Moya, Quast, Robert B., Robb, Nicole C., Sattler, Michael, Schlierf, Michael, Schneider, Jonathan, Schroeder, Tim, Sefer, Anna, Tan, Piau Siong, Thurn, Johann, Tinnefeld, Philip, van Noort, John, Weiss, Shimon, Wendler, Nicolas, Zijlstra, Niels, Barth, Anders, Seidel, Claus A. M., Lamb, Don C., Cordes, Thorben |
| المصدر: | Nature methods, vol 20, iss 4 |
| بيانات النشر: | Springer Science and Business Media LLC, 2023. |
| سنة النشر: | 2023 |
| مصطلحات موضوعية: | Technology, Molecular Conformation, Reproducibility of Results, Proteins, Cell Biology, Biological Sciences, Medical and Health Sciences, Biochemistry, Affordable and Clean Energy, Fluorescence Resonance Energy Transfer, Laboratories, Molecular Biology, Developmental Biology, Biotechnology |
| الوصف: | Funder: Ludwig Maximilians University Munich | Center for NanoScience, Ludwig-Maximilians-Universität Mnchen (CeNS, LMU); doi: https://doi.org/10.13039/501100007153 Funder: Alexander von Humboldt-Stiftung (Alexander von Humboldt Foundation); doi: https://doi.org/10.13039/100005156 Single-molecule Förster-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 Å and accuracy of ≤5 Å. 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. |
| وصف الملف: | application/pdf; text/xml; application/zip |
| DOI: | 10.17863/cam.96236 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::590d2e38ad76d3e86c3dfc774929abee |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....590d2e38ad76d3e86c3dfc774929abee |
| قاعدة البيانات: | OpenAIRE |
| DOI: | 10.17863/cam.96236 |
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