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Analysis of self-assembly of S-layer protein slp-B53 from Lysinibacillus sphaericus

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Abstract

The formation of stable and functional surface layers (S-layers) via self-assembly of surface-layer proteins on the cell surface is a dynamic and complex process. S-layers facilitate a number of important biological functions, e.g., providing protection and mediating selective exchange of molecules and thereby functioning as molecular sieves. Furthermore, S-layers selectively bind several metal ions including uranium, palladium, gold, and europium, some of them with high affinity. Most current research on surface layers focuses on investigating crystalline arrays of protein subunits in Archaea and bacteria. In this work, several complementary analytical techniques and methods have been applied to examine structure–function relationships and dynamics for assembly of S-layer protein slp-B53 from Lysinibacillus sphaericus: (1) The secondary structure of the S-layer protein was analyzed by circular dichroism spectroscopy; (2) Small-angle X-ray scattering was applied to gain insights into the three-dimensional structure in solution; (3) The interaction with bivalent cations was followed by differential scanning calorimetry; (4) The dynamics and time-dependent assembly of S-layers were followed by applying dynamic light scattering; (5) The two-dimensional structure of the paracrystalline S-layer lattice was examined by atomic force microscopy. The data obtained provide essential structural insights into the mechanism of S-layer self-assembly, particularly with respect to binding of bivalent cations, i.e., Mg2+ and Ca2+. Furthermore, the results obtained highlight potential applications of S-layers in the fields of micromaterials and nanobiotechnology by providing engineered or individual symmetric thin protein layers, e.g., for protective, antimicrobial, or otherwise functionalized surfaces.

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Acknowledgments

The work was supported by grants from the BMBF Röntgen–Ångström Cluster Project (RAC) under project number 05K12GU3 and by the DFG–Excellence Cluster CUI (Centre for Ultra-Fast Imaging) and Hamburg Ministry of Science and Research via the graduate school DELIGRAH. Jun Liu was assisted by the China Scholarship Council (CSC) via Grant No. 2010629147 and by the Technology Foundation for Selected Overseas Chinese Scholar, Ministry of Personnel of China. Sven Falke, Christian Betzel, and Dmitri Svergun further acknowledge support from a BMBF grant, referring to project number 05K13GU2.

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    Authors and Affiliations

    1. Laboratory for Structural Biology of Infection and Inflammation, Institute of Biochemistry and Molecular Biology, University of Hamburg, Martin-Luther-King Platz 6, 20146, Hamburg, Germany

      Jun Liu, Sven Falke, Dominik Oberthuer & Christian Betzel

    2. Bioengineering Faculty, Sichuan University of Science and Engineering, Huixing Rd., Xueyuan Street 180, Zigong, 643000, Sichuan, China

      Jun Liu

    3. Center for Free-Electron Laser Science (CFEL), DESY, Notkestr. 85, 22607, Hamburg, Germany

      Dominik Oberthuer

    4. EMBL Hamburg, European Molecular Biology Laboratory, Notkestr. 85, 22607, Hamburg, Germany

      Alexey Kikhney & Dmitri Svergun

    5. Institute of Resource Ecology, Helmholtz-Zentrum Dresden-Rossendorf, Bautzner Landstr. 400, 01328, Dresden, Germany

      Bjoern Drobot, Karim Fahmy & Johannes Raff

    6. Helmholtz Institute Freiberg for Resource Technology, Helmholtz-Zentrum Dresden-Rossendorf, Bautzner Landstr. 400, 01328, Dresden, Germany

      Tobias Guenther & Johannes Raff

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    Correspondence to Johannes Raff.

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    J. Liu and S. Falke contributed equally to this work.

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    Liu, J., Falke, S., Drobot, B. et al. Analysis of self-assembly of S-layer protein slp-B53 from Lysinibacillus sphaericus . Eur Biophys J 46, 77–89 (2017). https://doi.org/10.1007/s00249-016-1139-9

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