A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport
Abstract
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.
- Authors:
-
- 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
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Oak Ridge Leadership Computing Facility (OLCF); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1489406
- Grant/Contract Number:
- AC05-00OR22725; AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES
Citation Formats
Terry, Daniel S., Kolster, Rachel A., Quick, Matthias, LeVine, Michael V., Khelashvili, George, Zhou, Zhou, Weinstein, Harel, Javitch, Jonathan A., and Blanchard, Scott C. A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport. United States: N. p., 2018.
Web. doi:10.1038/s41467-017-02202-y.
Terry, Daniel S., Kolster, Rachel A., Quick, Matthias, LeVine, Michael V., Khelashvili, George, Zhou, Zhou, Weinstein, Harel, Javitch, Jonathan A., & Blanchard, Scott C. A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport. United States. https://doi.org/10.1038/s41467-017-02202-y
Terry, Daniel S., Kolster, Rachel A., Quick, Matthias, LeVine, Michael V., Khelashvili, George, Zhou, Zhou, Weinstein, Harel, Javitch, Jonathan A., and Blanchard, Scott C. Mon .
"A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport". United States. https://doi.org/10.1038/s41467-017-02202-y. https://www.osti.gov/servlets/purl/1489406.
@article{osti_1489406,
title = {A partially-open inward-facing intermediate conformation of LeuT is associated with Na+ release and substrate transport},
author = {Terry, Daniel S. and Kolster, Rachel A. and Quick, Matthias and LeVine, Michael V. and Khelashvili, George and Zhou, Zhou and Weinstein, Harel and Javitch, Jonathan A. and Blanchard, Scott C.},
abstractNote = {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.},
doi = {10.1038/s41467-017-02202-y},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {Mon Jan 15 00:00:00 EST 2018},
month = {Mon Jan 15 00:00:00 EST 2018}
}
Web of Science
Figures / Tables:
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