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The dataset also contains the cyro-EM maps and the low-pass-filtered cryo-EM maps.\n<br><br>\nBacteria utilize various strategies to prevent internal dehydration during hypertonic stress. A common approach to countering the effects of the stress is to import compatible solutes such as glycine betaine, leading to simultaneous passive water fluxes following the osmotic gradient. OpuA from Lactococcus lactis is a type I ABC-importer that uses two substrate-binding domains (SBDs) to capture extracellular glycine betaine and deliver the substrate to the transmembrane domains for subsequent transport. OpuA senses osmotic stress via changes in the internal ionic strength and is furthermore regulated by the 2nd messenger cyclic-di-AMP. We now show, by means of solution-based single-molecule FRET and analysis with multi-parameter photon-by-photon hidden Markov modeling, that the SBDs interact in an ionic strength-dependent manner. The smFRET data are in accordance with the apparent cooperativity in transport and supported by new cryo-EM data of OpuA. We propose that physical interactions between SBDs and cooperativity in substrate delivery could be more widespread than recognized thus far."}}]},{"typeName":"subject","multiple":true,"typeClass":"controlledVocabulary","value":["Medicine, Health and Life Sciences"]},{"typeName":"keyword","multiple":true,"typeClass":"compound","value":[{"keywordValue":{"typeName":"keywordValue","multiple":false,"typeClass":"primitive","value":"single-molecule FRET"}},{"keywordValue":{"typeName":"keywordValue","multiple":false,"typeClass":"primitive","value":"cryo-EM"}},{"keywordValue":{"typeName":"keywordValue","multiple":false,"typeClass":"primitive","value":"photon-by-photon hidden Markov modeling"}},{"keywordValue":{"typeName":"keywordValue","multiple":false,"typeClass":"primitive","value":"transient protein-protein 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