Single-molecule studies of conformational states and dynamics in the ABC importer OpuA

Tassis, Konstantinos; Vietrov, Ruslan; Koning, Matthijs de; Boer, Marijn de; Gouridis, Giorgos and Cordes, Thorben (2021): Single-molecule studies of conformational states and dynamics in the ABC importer OpuA. In: FEBS letters, Vol. 595, No. 6: pp. 717-734

Abstract

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.

• Item Type: Journal article
• EU Funded Grant Agreement Number: 638536
• EU Projects: Horizon 2020 > ERC Grants > ERC Starting Grant > ERC Grant 638536: SM-IMPORT - Substrate import at work: single-molecule studies of ABC transporters
• Faculties: Biology > Department Biology I
• Subjects: 500 Science > 570 Life sciences; biology
• Language: English