In the extremophile bacterium Deinococcus radiodurans, the outermost surface layer is tightly connected with the rest of the cell wall. This integrated organization provides a compact structure that shields the bacterium against environmental stresses. The fundamental unit of this surface layer (S-layer) is the S-layer deinoxanthin-binding complex (SDBC), which binds the carotenoid deinoxanthin and provides both, thermostability and UV radiation resistance. However, the structural organization of the SDBC awaits elucidation. Here, we report the isolation of the SDBC with a gentle procedure consisting of lysozyme treatment and solubilization with the nonionic detergent n-dodecyl-β-d-maltoside, which preserved both hydrophilic and hydrophobic components of the SDBC and allows the retention of several minor subunits. As observed by low-resolution single-particle analysis, we show that the complex possesses a porin-like structural organization, but is larger than other known porins. We also noted that the main SDBC component, the protein DR_2577, shares regions of similarity with known porins. Moreover, results from electrophysiological assays with membrane-reconstituted SDBC disclosed that it is a nonselective channel that has some peculiar gating properties, but also exhibits behavior typically observed in pore-forming proteins, such as porins and ionic transporters. The functional properties of this system and its porin-like organization provide information critical for understanding ion permeability through the outer cell surface of S-layer-carrying bacterial species.

Department of Life Sciences and Chemistry Jacobs University Bremen 28759 Bremen Germany

Department of Physics and IOM CNR University of Cagliari 09042 Monserrato Italy

Department of Plant Physiology Warsaw University of Life Sciences SGGW Nowoursynowska Str 159 02776 Warsaw Poland

Institute of Biology and Medical Genetics 1st Faculty of Medicine Charles University Prague 128 00 Czech Republic

Laboratory of Plant Physiology and Photobiology Department of Life and Environmental Sciences University of Cagliari 5 le S Ignazio da Laconi 13 09123 Cagliari Italy

Leibniz Institute on Ageing Fritz Lipmann Institute Beutenbergstrasse 11 07745 Jena Germany; The Francis Crick Institute 1 Midland Road NW1 1AT London United Kingdom

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The cryo-EM structure of the S-layer deinoxanthin-binding complex of Deinococcus radiodurans informs properties of its environmental interactions

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PDB
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