Noncovalent Functionalization of Carbon Substrates with Hydrogels Improves Structural Analysis of Vitrified Proteins by Electron Cryo-Microscopy
| Autor: | Daniel Rhinow, Bonnie J. Murphy, Julian Scherr, Alexander Neuhaus, Niklas Klusch, Volker Zickermann, Werner Kühlbrandt, Kristian Parey, Andreas Terfort |
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| Rok vydání: | 2019 |
| Předmět: |
Glycerol
Materials science Cryo-electron microscopy Polymers Detergents General Physics and Astronomy chemistry.chemical_element Carbon nanotube law.invention law General Materials Science Nanotubes Pyrenes Protein Stability Cryoelectron Microscopy General Engineering Membrane Proteins Hydrogels Vitrification Amorphous carbon Chemical engineering chemistry Membrane protein complex Self-healing hydrogels Surface modification Self-assembly Carbon |
| Zdroj: | ACS nano. 13(6) |
| ISSN: | 1936-086X |
| Popis: | In electron cryo-microscopy, structure determination of protein molecules is frequently hampered by adsorption of the particles to the support film material, typically amorphous carbon. Here, we report that pyrene derivatives with one or two polyglycerol (PG) side chains bind to the amorphous carbon films, forming a biorepulsive hydrogel layer so that the number of protein particles in the vitreous ice drastically increases. This approach could be extended by adding a hydrogel-functionalized carbon nanotube network (HyCaNet, the hydrogel again being formed from the PG-pyrene derivatives), which stabilized the protein-containing thin ice films during imaging with the electron beam. The stabilization resulted in reduced particle motion by up to 70%. These substrates were instrumental for determining the structure of a large membrane protein complex. |
| Databáze: | OpenAIRE |
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