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Single‐molecule studies of conformational states and dynamics in the ABC importer OpuA
Author(s) -
Tassis Konstantinos,
Vietrov Ruslan,
Koning Matthijs,
Boer Marijn,
Gouridis Giorgos,
Cordes Thorben
Publication year - 2021
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.14026
Subject(s) - single molecule fret , atp binding cassette transporter , förster resonance energy transfer , conformational change , biophysics , chemistry , transmembrane domain , biology , biochemistry , transporter , membrane , physics , fluorescence , quantum mechanics , gene
The current model of active transport via ABC importers is mostly based on structural, biochemical and genetic data. We here establish single‐molecule Förster 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.