Lerner Eitan, Barth Anders, Hendrix Jelle, Ambrose Benjamin, Birkedal Victoria, Blanchard Scott C, Börner Richard, Sung Chung Hoi, Cordes Thorben, Craggs Timothy D, Deniz Ashok A, Diao Jiajie, Fei Jingyi, Gonzalez Ruben L, Gopich Irina V, Ha Taekjip, Hanke Christian A, Haran Gilad, Hatzakis Nikos S, Hohng Sungchul, Hong Seok-Cheol, Hugel Thorsten, Ingargiola Antonino, Joo Chirlmin, Kapanidis Achillefs N, Kim Harold D, Laurence Ted, Lee Nam Ki, Lee Tae-Hee, Lemke Edward A, Margeat Emmanuel, Michaelis Jens, Michalet Xavier, Myong Sua, Nettels Daniel, Peulen Thomas-Otavio, Ploetz Evelyn, Razvag Yair, Robb Nicole C, Schuler Benjamin, Soleimaninejad Hamid, Tang Chun, Vafabakhsh Reza, Lamb Don C, Seidel Claus Am, Weiss Shimon
Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, and The Center for Nanoscience and Nanotechnology, Faculty of Mathematics & Science, The Edmond J. Safra Campus, The Hebrew University of Jerusalem, Jerusalem, Israel.
Lehrstuhl für Molekulare Physikalische Chemie, Heinrich-Heine-Universität, Düsseldorf, Germany.
Elife. 2021 Mar 29;10:e60416. doi: 10.7554/eLife.60416.
Single-molecule FRET (smFRET) has become a mainstream technique for studying biomolecular structural dynamics. The rapid and wide adoption of smFRET experiments by an ever-increasing number of groups has generated significant progress in sample preparation, measurement procedures, data analysis, algorithms and documentation. Several labs that employ smFRET approaches have joined forces to inform the smFRET community about streamlining how to perform experiments and analyze results for obtaining quantitative information on biomolecular structure and dynamics. The recent efforts include blind tests to assess the accuracy and the precision of smFRET experiments among different labs using various procedures. These multi-lab studies have led to the development of smFRET procedures and documentation, which are important when submitting entries into the archiving system for integrative structure models, PDB-Dev. This position paper describes the current 'state of the art' from different perspectives, points to unresolved methodological issues for quantitative structural studies, provides a set of 'soft recommendations' about which an emerging consensus exists, and lists openly available resources for newcomers and seasoned practitioners. To make further progress, we strongly encourage 'open science' practices.
单分子荧光共振能量转移（smFRET）已成为研究生物分子结构动力学的主流技术。越来越多的研究团队迅速且广泛地采用smFRET实验，这在样品制备、测量程序、数据分析、算法及文档记录等方面都取得了显著进展。几个采用smFRET方法的实验室联合起来，向smFRET领域的同行介绍如何简化实验操作及分析结果，以便获取生物分子结构和动力学的定量信息。近期的工作包括进行盲测，以评估不同实验室使用各种程序开展smFRET实验的准确性和精密度。这些多实验室研究推动了smFRET程序及文档记录的发展，这对于向整合结构模型存档系统PDB-Dev提交数据条目而言至关重要。本立场文件从不同角度描述了当前的“技术现状”，指出了定量结构研究中尚未解决的方法学问题，提供了一组已形成初步共识的“软性建议”，并为新手和经验丰富的从业者列出了公开可用的资源。为取得进一步进展，我们大力鼓励“开放科学”实践。
