Journal Article

Conformational and dynamic plasticity in substrate-binding proteins underlies selective transport in ABC importers

de Boer, Marijn, Husada, Florence, McDevitt, Christopher A, Eleftheriadis, Nikolaos, Vietrov, Ruslan, Begg, Stephanie L, Schuurman-Wolters, Gea K, Gouridis, Giorgos, Cordes, Thorben, Poolman, Bert

2019

Overview

Substrate-binding proteins (SBPs) are associated with ATP-binding cassette importers and switch from an open to a closed conformation upon substrate binding, providing specificity for transport. We investigated the effect of substrates on the conformational dynamics of six SBPs and the impact on transport. Using single-molecule FRET, we reveal an unrecognized diversity of plasticity in SBPs. We show that a unique closed SBP conformation does not exist for transported substrates. Instead, SBPs sample a range of conformations that activate transport. Certain non-transported ligands leave the structure largely unaltered or trigger a conformation distinct from that of transported substrates. Intriguingly, in some cases, similar SBP conformations are formed by both transported and non-transported ligands. In this case, the inability for transport arises from slow opening of the SBP or the selectivity provided by the translocator. Our results reveal the complex interplay between ligand-SBP interactions, SBP conformational dynamics and substrate transport.

• Publisher: eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
• Subject: ABC transporter / ATP-Binding Cassette Transporters - chemistry / ATP-Binding Cassette Transporters - metabolism / Biochemistry and Chemical Biology / biophysics / Conformation / Fluorescence Resonance Energy Transfer / Immunology / Kinetics / Ligands / membrane transport / NMR / Nuclear magnetic resonance / Plasticity / Protein Binding / Protein Conformation / Proteins / Single Molecule Imaging / single-molecule / smFRET / Structural Biology and Molecular Biophysics / Substrate Specificity / substrate-binding protein