Structural Changes in Tubulin Sheets Caused by Immobilization on Solid Supports
| Autor: | William B. Jobs, Evan C. Sheffield, Elysa J. Kliman, Visham Appadoo, Zachary J. Donhauser |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Materials science
General Chemical Engineering 02 engineering and technology macromolecular substances Article law.invention lcsh:Chemistry 03 medical and health sciences Scanning probe microscopy Protein structure law Microtubule Pyrolytic carbon 030304 developmental biology 0303 health sciences biology General Chemistry 021001 nanoscience & nanotechnology Tubulin lcsh:QD1-999 biology.protein Biophysics Mica Electron microscope 0210 nano-technology Protein crystallization |
| Zdroj: | ACS Omega ACS Omega, Vol 3, Iss 12, Pp 18196-18202 (2018) |
| ISSN: | 2470-1343 |
| Popis: | Open in a separate window In the presence of zinc, the protein tubulin assembles into two-dimensional sheets that are a useful model system for the study of both tubulin and microtubule structure. Tubulin sheets present an ideal protein structure for study with atomic force microscopy because they contain a two-dimensional crystalline protein lattice and retain many of the structural features of tubulin and microtubules. However, high-resolution imaging requires nonperturbative immobilization onto an appropriate imaging substrate. In this report, several substrates commonly used for scanning probe microscopy are evaluated for their ability to effectively immobilize tubulin sheets: mica, gold, highly ordered pyrolytic graphite, and carbon-coated electron microscopy grids. We hypothesize that the different intermolecular interactions presented by these substrates will affect the morphology of adsorbed tubulin sheets as well as the amount of other contaminating adsorbates. Tubulin sheets were successfully imaged on all of these substrates and structural characterization is reported. The most consistent results were obtained on carbon-coated electron microscopy grids, which preserved fine structural features of the sheets and had the least amount of contamination from the adsorption of unpolymerized tubulin. Images of tubulin sheets obtained with atomic force microscopy also compare favorably with published electron micrographs of sheets produced using similar procedures. This work demonstrates the importance of assessing substrate effects when studying two-dimensional protein crystals and identifies suitable substrates for immobilizing tubulin sheets. |
| Databáze: | OpenAIRE |
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