Abstract
N-Methyl-D-aspartate (NMDA) receptors are the main calcium-permeable excitatory receptors in the mammalian central nervous system. The NMDA receptor gating is complex, exhibiting multiple closed, open, and desensitized states; however, central questions regarding the conformations and energetics of the transmembrane domains as they relate to the gating states are still unanswered. Here, using single-molecule Förster resonance energy transfer (smFRET), we map the energy landscape of the first transmembrane segment of the Rattus norvegicus NMDA receptor under resting and various liganded conditions. These results show kinetically and structurally distinct changes associated with apo, agonist-bound, and inhibited receptors linked by a linear mechanism of gating at this site. Furthermore, the smFRET data suggest that allosteric inhibition by zinc occurs by an uncoupling of the agonist-induced changes at the extracellular domains from the gating motions leading to an apo-like state, while dizocilpine, a pore blocker, stabilizes multiple closely packed transmembrane states.
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Acknowledgements
Methods and additional data can be found in the supplementary materials. This project was supported by NIH grant R35GM122528 to V.J., K99NS094761to D.M.M., American Heart Association Fellowship 16POST30030007 to S.A.S., Schissler Foundation Fellowship to D.M.D., and Welch Foundation Grant C-1787 to C.F.L.
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D.M.D. performed the mutations and prepared the labeled proteins, analyzed the smFRET data, and contributed to designing the research, interpreting the results, and writing the manuscript. S.C. performed the smFRET measurements, analyzed the smFRET data, and contributed to designing the research, interpreting the results, and writing the manuscript. D.M.M. and S.A.S. performed the electrophysiology. C.F. and L.D.C.B. analyzed the smFRET data. C.F.L., and V.J. analyzed the smFRET data and contributed to designing the research, interpreting the results, and writing the manuscript.
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Dolino, D., Chatterjee, S., MacLean, D. et al. The structure–energy landscape of NMDA receptor gating. Nat Chem Biol 13, 1232–1238 (2017). https://doi.org/10.1038/nchembio.2487
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DOI: https://doi.org/10.1038/nchembio.2487
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