Flexible, symmetry-directed approach to assembling protein cages
| Autor: | Joseph D. Eschweiler, Min Su, James C.A. Bardwell, Kelsey Diffley, Aaron Sciore, Philipp Koldewey, Brian M. Linhares, E. Neil G. Marsh, Brandon T. Ruotolo, Georgios Skiniotis |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Models Molecular Protein Folding Protein subunit Protein domain Protein design Sequence (biology) Biology 010402 general chemistry 01 natural sciences Mass Spectrometry Protein Structure Secondary 03 medical and health sciences Microscopy Electron Transmission Amino Acid Sequence Peptide sequence Multidisciplinary Cryoelectron Microscopy Proteins Biological Sciences Negative stain 0104 chemical sciences Crystallography 030104 developmental biology Biophysics Protein folding Protein Multimerization Octamer Transcription Factor-2 Linker Octamer Transcription Factor-3 |
| Popis: | The assembly of individual protein subunits into large-scale symmetrical structures is widespread in nature and confers new biological properties. Engineered protein assemblies have potential applications in nanotechnology and medicine; however, a major challenge in engineering assemblies de novo has been to design interactions between the protein subunits so that they specifically assemble into the desired structure. Here we demonstrate a simple, generalizable approach to assemble proteins into cage-like structures that uses short de novo designed coiled-coil domains to mediate assembly. We assembled eight copies of a C3-symmetric trimeric esterase into a well-defined octahedral protein cage by appending a C4-symmetric coiled-coil domain to the protein through a short, flexible linker sequence, with the approximate length of the linker sequence determined by computational modeling. The structure of the cage was verified using a combination of analytical ultracentrifugation, native electrospray mass spectrometry, and negative stain and cryoelectron microscopy. For the protein cage to assemble correctly, it was necessary to optimize the length of the linker sequence. This observation suggests that flexibility between the two protein domains is important to allow the protein subunits sufficient freedom to assemble into the geometry specified by the combination of C4 and C3 symmetry elements. Because this approach is inherently modular and places minimal requirements on the structural features of the protein building blocks, it could be extended to assemble a wide variety of proteins into structures with different symmetries. |
| Databáze: | OpenAIRE |
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