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Structural landscape of isolated agonist-binding domains from single AMPA receptors

Nature Chemical Biology volume 7, pages 168–173 (2011)Cite this article

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Abstract

AMPA receptors mediate fast excitatory neurotransmission by converting chemical signals into electrical signals, and thus it is important to understand the relationship between their chemical biology and their function. We used single-molecule fluorescence resonance energy transfer to examine the conformations explored by the agonist-binding domain of the AMPA receptor for wild-type and T686S mutant proteins. Each form of the agonist binding domain showed a dynamic, multistate sequential equilibrium, which could be identified only using wavelet shrinkage, a signal processing technique that removes experimental shot noise. These results illustrate that the extent of activation depends not on a rigid closed cleft but instead on the probability that a given subunit will occupy a closed-cleft conformation, which in turn is determined not only by the lowest energy state but also by the range of states that the protein explores.

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Figure 1: Crystal structure of GluA2-ABD.
Figure 2: Single-molecule smFRET before and after denoising.
Figure 3: Ensemble smFRET histograms.
Figure 4: Dwell-time histograms for all of the observed transitions between the adjacent four states identified from the glutamate-bound GluA2-ABD form.
Figure 5: Denoised ensemble smFRET histograms.
Figure 6: Average smFRET efficiency autocorrelation.

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Acknowledgements

C.F.L. thanks the Norman Hackerman Welch Young Investigator Program at Rice University. We acknowledge the Donors of the American Chemical Society Petroleum Research Fund for partial support of this research (to C.F.L.). This work was supported by US National Institutes of Health Grant R01GM073102 (to V.J.) and American Heart Association Grant 0855081F (V.J.).We also thank H. Yang and R. Goldsmith for suggestions.

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Authors and Affiliations

  1. Department of Chemistry, Rice University, Houston, Texas, USA

    Christy F Landes, J Nick Taylor & Ferandre Salatan

  2. Department of Biochemistry and Molecular Biology, Center for Membrane Biology, Graduate School of Biomedical Sciences, University of Texas Health Science Center, Houston, Texas, USA

    Anu Rambhadran & Vasanthi Jayaraman

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Contributions

C.F.L. and V.J. directed the research and wrote the manuscript. A.R. prepared and purified the protein samples and performed single-molecule experiments. J.N.T. analyzed the single-molecule data and prepared figures. F.S. performed single-molecule experiments and analyzed data.

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Correspondence to Christy F Landes or Vasanthi Jayaraman.

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Landes, C., Rambhadran, A., Taylor, J. et al. Structural landscape of isolated agonist-binding domains from single AMPA receptors. Nat Chem Biol 7, 168–173 (2011). https://doi.org/10.1038/nchembio.523

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