Design of biologically active binary protein 2D materials
| Autor: | Hannele Ruohola-Baker, Jiajun Chen, Emmanuel Derivery, Clemens F. Kaminski, Stephen C. Blacklow, William Sheffler, Ioanna Mela, Justin M. Kollman, Matthew C. Johnson, David Baker, Andrew A. Drabek, Fang Jiao, Ariel J. Ben-Sasson, Joseph L. Watson, Logeshwaran Somasundaram, James J. De Yoreo, Alice Bittleston, Sanchez M. Jarrett, Justin Decarreau, Greg L. Hura |
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| Rok vydání: | 2019 |
| Předmět: |
Streptavidin
Models Molecular Materials science Cell Survival Protein Array Analysis Nanotechnology 02 engineering and technology Dihedral angle In Vitro Techniques Endocytosis Ligands Microscopy Atomic Force Protein Engineering Article 03 medical and health sciences Synthetic biology chemistry.chemical_compound Mice Nanocages Cell surface receptor Escherichia coli Animals 030304 developmental biology 0303 health sciences Multidisciplinary Ligand Computational Biology Proteins Protein engineering 3T3 Cells Cell Biology 021001 nanoscience & nanotechnology Kinetics Membrane chemistry Drug Design Protein microarray Biophysics Synthetic Biology Receptor clustering 0210 nano-technology Protein crystallization |
| Zdroj: | Nature |
| ISSN: | 1476-4687 |
| Popis: | Ordered two-dimensional arrays such as S-layers1,2 and designed analogues3–5 have intrigued bioengineers,6,7 but with the exception of a single lattice formed with flexible linkers,8 they are constituted from just one protein component. For modulating assembly dynamics and incorporating more complex functionality, materials composed of two components would have considerable advantages.9–12 Here we describe a computational method to generate co-assembling binary layers by designing rigid interfaces between pairs of dihedral protein building-blocks, and use it to design a p6m lattice. The designed array components are soluble at mM concentrations, but when combined at nM concentrations, rapidly assemble into nearly crystalline micrometer-scale arrays nearly identical (based on TEM and SAXS) to the computational design model in vitro and in cells without the need for a two-dimensional support. Because the material is designed from the ground up, the components can be readily functionalized, and their symmetry reconfigured, enabling formation of ligand arrays with distinguishable surfaces which we demonstrate can drive extensive receptor clustering, downstream protein recruitment, and signaling. Using AFM on supported bilayers and quantitative microscopy on living cells, we show that arrays assembled on membranes have component stoichiometry and structure similar to arrays formed in vitro, and thus that our material can impose order onto fundamentally disordered substrates like cell membranes. In sharp contrast to previously characterized cell surface receptor binding assemblies such as antibodies and nanocages, which are rapidly endocytosed, we find that large arrays assembled at the cell surface suppress endocytosis in a tunable manner, with potential therapeutic relevance for extending receptor engagement and immune evasion. Our work paves the way towards a synthetic cell biology, where a new generation of multi-protein macroscale materials is designed to modulate cell responses and reshape synthetic and living systems. One Sentence Summary: Design of a two component protein array enables robust formation of complex large scale ordered biologically active materials |
| Databáze: | OpenAIRE |
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