A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport

Terry, Daniel S.; Kolster, Rachel A.; Quick, Matthias; LeVine, Michael V.; Khelashvili, George; Zhou, Zhou; Weinstein, Harel; Javitch, Jonathan A.; Blanchard, Scott C.

doi:10.1038/s41467-017-02202-y

A partially-open inward-facing intermediate conformation of LeuT is associated with Na⁺ release and substrate transport

Journal Article · Mon Jan 15 00:00:00 EST 2018 · Nature Communications

DOI:https://doi.org/10.1038/s41467-017-02202-y· OSTI ID:1489406

^[1]; Kolster, Rachel A. ^[2]; Quick, Matthias ^[2]; LeVine, Michael V. ^[3]; Khelashvili, George ^[3]; Zhou, Zhou ^[1]; ^[3]; Javitch, Jonathan A. ^[4]; Blanchard, Scott C. ^[1]

Weill Cornell Medicine, New York, NY (United States). Dept. of Physiology and Biophysics
Columbia Univ. College of Physicians and Surgeons and New York State Psychiatric Inst, New York, NY (United States). Dept. of Psychiatry
Weill Cornell Medicine, New York, NY (United States). Dept. of Physiology and Biophysics and HRH Prince Alwaleed Bin Talal Bin Abdulazaz Alsaud Inst. for Computational Biomedicine
Columbia Univ. College of Physicians and Surgeons and New York State Psychiatric Inst, New York, NY (United States). Dept. of Psychiatry and Dept. of Pharmacology

Here, neurotransmitter:sodium symporters (NSS), targets of antidepressants and psychostimulants, clear neurotransmitters from the synaptic cleft through sodium (Na⁺)-coupled transport. Substrate and Na⁺ are thought to be transported from the extracellular to intracellular space through an alternating access mechanism by coordinated conformational rearrangements in the symporter that alternately expose the binding sites to each side of the membrane. However, the mechanism by which the binding of ligands coordinates conformational changes occurring on opposite sides of the membrane is not well understood. Here, we report the use of single-molecule fluorescence resonance energy transfer (smFRET) techniques to image transitions between distinct conformational states on both the extracellular and intracellular sides of the prokaryotic NSS LeuT, including partially open intermediates associated with transport activity. The nature and functional context of these hitherto unidentified intermediate states shed new light on the allosteric mechanism that couples substrate and Na⁺ symport by the NSS family through conformational dynamics.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center; Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231; AC05-00OR22725

OSTI ID:: 1489406

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 9; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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