The goal of this project is to use new advances in cryo-electron microscopy (cryo-EM) technology to elucidate the structures of, at high-resolution, a range of functional states of ionotropic glutamate receptor (iGluR) ion channels, and to use complementary functional and biochemical assays to validate our findings and hypotheses driven by our solved cryo-EM models. iGluRs are responsible for the majority of excitatory neurotransmission, and are implicated in a wide range of pathologies, from neurodegenerative diseases and psychiatric disorders, to epilepsy and stroke-induced trauma. Current design of therapeutics targeting the range of iGluR functional states that dictate these different disease states is hindered by a lack of information describing iGluRs outside of a closed-channel state. This research will directly address the need for new views into these functional states by using cryo-EM based methods, which will allow crystallographic barriers in analyzing iGluR states to be overcome. Using state-of-the-art cryo-EM techniques, high-resolution cryo-EM of iGluRs will be established. With fluorescence-based construct screening strategies, new targets for isolating separate structural states in iGluR function will be identified. Namely, how auxiliary proteins interact with/regulate iGluRs and iGluR gating will be targeted. After cryo-EM analysis, data will be probed using advanced electrophysiological techniques and biochemical interrogation. The methods used in this project will serve as a foundation for investigating functional states across ion channel families, and may provide a much-needed template for structure-based drug design of new molecules targeting iGluR states.
The proposed research targets the elucidation of three-dimensional structures of various functional states in a group of ion channels called ionotropic glutamate receptors (iGluRs). These structures will provide highly detailed information to help better understand how iGluRs function across various disease-related states, such as in neurodegenerative diseases and psychiatric disorders. The outcomes of this project will provide a foundation, from a structural biology perspective, for novel design of therapeutics targeting an array of iGluR functional states across neurological disorders.
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| Twomey, Edward C; Yelshanskaya, Maria V; Vassilevski, Alexander A et al. (2018) Mechanisms of Channel Block in Calcium-Permeable AMPA Receptors. Neuron 99:956-968.e4 |
| McGoldrick, Luke L; Singh, Appu K; Saotome, Kei et al. (2018) Opening of the human epithelial calcium channel TRPV6. Nature 553:233-237 |
| Singh, Appu K; McGoldrick, Luke L; Twomey, Edward C et al. (2018) Mechanism of calmodulin inactivation of the calcium-selective TRP channel TRPV6. Sci Adv 4:eaau6088 |
| Twomey, Edward C; Yelshanskaya, Maria V; Grassucci, Robert A et al. (2017) Channel opening and gating mechanism in AMPA-subtype glutamate receptors. Nature 549:60-65 |
| Twomey, Edward C; Yelshanskaya, Maria V; Grassucci, Robert A et al. (2017) Structural Bases of Desensitization in AMPA Receptor-Auxiliary Subunit Complexes. Neuron 94:569-580.e5 |
| Twomey, Edward C; Yelshanskaya, Maria V; Grassucci, Robert A et al. (2016) Elucidation of AMPA receptor-stargazin complexes by cryo-electron microscopy. Science 353:83-6 |
