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Instruments / Technical Area	Publication Details	Published
	Structure and dynamics of the E. coli chemotaxis core signaling complex by cryo-electron tomography and molecular simulations C. Keith Cassidy , Benjamin A. Himes , Dapeng Sun , Jun Ma , Gongpu Zhao , John S. Parkinson , Phillip J. Stansfeld , Zaida Luthey-Schulten , Peijun Zhang Communications Biology 3 DOI: 10.1038/s42003-019-0748-0 Open Access Show Abstract ▼ Abstract: To enable the processing of chemical gradients, chemotactic bacteria possess large arrays of transmembrane chemoreceptors, the histidine kinase CheA, and the adaptor protein CheW, organized as coupled core-signaling units (CSU). Despite decades of study, important questions surrounding the molecular mechanisms of sensory signal transduction remain unresolved, owing especially to the lack of a high-resolution CSU structure. Here, we use cryo-electron tomography and sub-tomogram averaging to determine a structure of the Escherichia coli CSU at sub-nanometer resolution. Based on our experimental data, we use molecular simulations to construct an atomistic model of the CSU, enabling a detailed characterization of CheA conformational dynamics in its native structural context. We identify multiple, distinct conformations of the critical P4 domain as well as asymmetries in the localization of the P3 bundle, offering several novel insights into the CheA signaling mechanism.	Dec 2020
Krios IV-Titan Krios IV at Diamond	Complete structure of the chemosensory array core signalling unit in an E. coli minicell strain Alister Burt , C. Keith Cassidy , Peter Ames , Maria Bacia-Verloop , Megghane Baulard , Karine Huard , Zaida Luthey-Schulten , Ambroise Desfosses , Phillip J. Stansfeld , William Margolin , John S. Parkinson , Irina Gutsche Nature Communications 11 DOI: 10.1038/s41467-020-14350-9 Diamond Proposal Number(s): [18112] Open Access Show Abstract ▼ Abstract: Motile bacteria sense chemical gradients with transmembrane receptors organised in supramolecular signalling arrays. Understanding stimulus detection and transmission at the molecular level requires precise structural characterisation of the array building block known as a core signalling unit. Here we introduce an Escherichia coli strain that forms small minicells possessing extended and highly ordered chemosensory arrays. We use cryo-electron tomography and subtomogram averaging to provide a three-dimensional map of a complete core signalling unit, with visible densities corresponding to the HAMP and periplasmic domains. This map, combined with previously determined high resolution structures and molecular dynamics simulations, yields a molecular model of the transmembrane core signalling unit and enables spatial localisation of its individual domains. Our work thus offers a solid structural basis for the interpretation of a wide range of existing data and the design of further experiments to elucidate signalling mechanisms within the core signalling unit and larger array.	Feb 2020
	CryoEM-based hybrid modeling approaches for structure determination C. Keith Cassidy , Benjamin A. Himes , Zaida Luthey-Schulten , Peijun Zhang Current Opinion In Microbiology 43, 14 - 23 DOI: 10.1016/j.mib.2017.10.002 Show Abstract ▼ Abstract: Recent advances in cryo-electron microscopy (cryoEM) have dramatically improved the resolutions at which vitrified biological specimens can be studied, revealing new structural and mechanistic insights over a broad range of spatial scales. Bolstered by these advances, much effort has been directed toward the development of hybrid modeling methodologies for the construction and refinement of high-fidelity atomistic models from cryoEM data. In this brief review, we will survey the key elements of cryoEM-based hybrid modeling, providing an overview of available computational tools and strategies as well as several recent applications.	Nov 2017

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