In Situ Structure of an Intact Lipopolysaccharide-Bound Bacterial Surface Layer.

Andriko von Kügelgen, Haiping Tang, Gail G Hardy, Danguole Kureisaite-Ciziene, Yves V Brun, Phillip J Stansfeld, Carol V Robinson, Tanmay A M Bharat

Cell 2020 Jan 23

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Most bacterial and all archaeal cells are encapsulated by a paracrystalline, protective, and cell-shape-determining proteinaceous surface layer (S-layer). On Gram-negative bacteria, S-layers are anchored to cells via lipopolysaccharide. Here, we report an electron cryomicroscopy structure of the Caulobacter crescentus S-layer bound to the O-antigen of lipopolysaccharide. Using native mass spectrometry and molecular dynamics simulations, we deduce the length of the O-antigen on cells and show how lipopolysaccharide binding and S-layer assembly is regulated by calcium. Finally, we present a near-atomic resolution in situ structure of the complete S-layer using cellular electron cryotomography, showing S-layer arrangement at the tip of the O-antigen. A complete atomic structure of the S-layer shows the power of cellular tomography for in situ structural biology and sheds light on a very abundant class of self-assembling molecules with important roles in prokaryotic physiology with marked potential for synthetic biology and surface-display applications. Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.

Citation

Andriko von Kügelgen, Haiping Tang, Gail G Hardy, Danguole Kureisaite-Ciziene, Yves V Brun, Phillip J Stansfeld, Carol V Robinson, Tanmay A M Bharat. In Situ Structure of an Intact Lipopolysaccharide-Bound Bacterial Surface Layer. Cell. 2020 Jan 23;180(2):348-358.e15