@article{facdbcd8f8104f0daff50eda397144d3,
title = "Single-molecule studies of conformational states and dynamics in the ABC importer OpuA",
abstract = "The current model of active transport via ABC importers is mostly based on structural, biochemical and genetic data. We here establish single-molecule F{\"o}rster resonance energy transfer (smFRET) assays to monitor the conformational states and heterogeneity of the osmoregulatory type I ABC importer OpuA from Lactococcus lactis. We present data probing both intradomain distances that elucidate conformational changes within the substrate-binding domain (SBD) OpuAC, and interdomain distances between SBDs or transmembrane domains. Using this methodology, we studied ligand-binding mechanisms, as well as ATP and glycine betaine dependences of conformational changes. Our work expands the scope of smFRET investigations towards a class of so far unstudied ABC importers, and paves the way for a full understanding of their transport cycle in the future.",
keywords = "ABC transporter, conformational dynamics, membrane transport, osmoregulation, single-molecule Foerster resonance energy transfer, substrate-binding domains",
author = "Konstantinos Tassis and Ruslan Vietrov and \{de Koning\}, Matthijs and \{de Boer\}, Marijn and Giorgos Gouridis and Thorben Cordes",
note = "Funding Information: This work was financed by an NWO Veni grant (722.012.012 to GG), an ERC Starting Grant (ERC‐STG 638536 – SM‐IMPORT to TC) and the Zernike Institute for Advanced Materials (support to GG and TC). GG also acknowledges an EMBO fellowship (long‐term fellowship ALF 47‐2012), and the Rega Foundation Postdoctoral Programme of KU Leuven. We thank J. Hammerl for preparation of Fig. 4 and C. Gebhardt for providing data analysis scripts. We thank B. Poolman for continuous support of the project, which included conceptional discussions on the structure and function of OpuA and access to materials and protocols. We finally thank G. K. Schuurman‐Wolters for experimental support, and B. Poolman and D. A. Griffith for critically reading the manuscript. Publisher Copyright: {\textcopyright} 2020 The Authors. FEBS Letters published by John Wiley \& Sons Ltd on behalf of Federation of European Biochemical Societies.",
year = "2021",
month = mar,
doi = "10.1002/1873-3468.14026",
language = "English",
volume = "595",
pages = "717--734",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Wiley",
number = "6",
}