X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release
| Autor: | Jonas S. Mortensen, Julie Winkel Missel, Jonathan A. Javitch, Ulrik Gether, George Khelashvili, Poul Nissen, Harel Weinstein, Matthias Quick, Scott C. Blanchard, Michael V. LeVine, Thomas Boesen, Pontus Gourdon, Daniel S. Terry, Kamil Gotfryd, Claus J. Loland |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Protein Conformation
Science Biophysics General Physics and Astronomy Context (language use) Molecular Dynamics Simulation Crystallography X-Ray Plasma Membrane Neurotransmitter Transport Proteins Biochemistry Article General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Molecular dynamics 0302 clinical medicine Protein structure Bacterial Proteins Leucine Binding site lcsh:Science 030304 developmental biology 0303 health sciences Binding Sites Multidisciplinary Bacteria Chemistry General Chemistry Computational biology and bioinformatics 3. Good health Aquifex Transmembrane domain Amino Acid Substitution Cytoplasm Symporter Mutagenesis Site-Directed lcsh:Q Structural biology 030217 neurology & neurosurgery Intracellular Neuroscience |
| Zdroj: | Gotfryd, K, Boesen, T, Mortensen, J S, Khelashvili, G, Quick, M, Terry, D S, Missel, J W, LeVine, M V, Gourdon, P, Blanchard, S C, Javitch, J A, Weinstein, H, Loland, C J, Nissen, P & Gether, U 2020, ' X-ray structure of LeuT in an inward-facing occluded conformation reveals mechanism of substrate release ', Nature Communications, vol. 11, no. 1, 1005 . https://doi.org/10.1038/s41467-020-14735-w Nature Communications, Vol 11, Iss 1, Pp 1-14 (2020) Nature Communications |
| DOI: | 10.1038/s41467-020-14735-w |
| Popis: | Neurotransmitter:sodium symporters (NSS) are conserved from bacteria to man and serve as targets for drugs, including antidepressants and psychostimulants. Here we report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation. To obtain this structure, we were guided by findings from single-molecule fluorescence spectroscopy and molecular dynamics simulations indicating that L-Phe binding and mutation of the conserved N-terminal Trp8 to Ala both promote an inward-facing state. Compared to the outward-facing occluded conformation, our structure reveals a major tilting of the cytoplasmic end of transmembrane segment (TM) 5, which, together with release of the N-terminus but without coupled movement of TM1, opens a wide cavity towards the second Na+ binding site. The structure of this key intermediate in the LeuT transport cycle, in the context of other NSS structures, leads to the proposal of an intracellular release mechanism of substrate and ions in NSS proteins. Neurotransmitter:sodium symporters (NSS) serve as targets for drugs including antidepressants and psychostimulants. Here authors report the X-ray structure of the prokaryotic NSS member, LeuT, in a Na+/substrate-bound, inward-facing occluded conformation which is a key intermediate in the LeuT transport cycle. |
| Databáze: | OpenAIRE |
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