A Versatile System for High-Throughput In Situ X-ray Screening and Data Collection of Soluble and Membrane-Protein Crystals
| Autor: | Aidin R. Balo, Oliver P. Ernst, Wei-Lin Ou, A. Kuo, Craig M. Ogata, Jana Broecker, David J. Kissick, Viviane Klingel |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
In situ 030103 biophysics Materials science biology X-ray Oxygen transport Bacteriorhodopsin Nanotechnology General Chemistry Haloquadratum walsbyi Condensed Matter Physics biology.organism_classification Article law.invention Crystal 03 medical and health sciences 030104 developmental biology law X-ray crystallography biology.protein General Materials Science Crystallization |
| Zdroj: | Crystal Growth & Design |
| ISSN: | 1528-7505 1528-7483 |
| Popis: | In recent years, in situ data collection has been a major focus of progress in protein crystallography. Here, we introduce the Mylar in situ method using Mylar-based sandwich plates that are inexpensive, easy to make and handle, and show significantly less background scattering than other setups. A variety of cognate holders for patches of Mylar in situ sandwich films corresponding to one or more wells makes the method robust and versatile, allows for storage and shipping of entire wells, and enables automated crystal imaging, screening, and goniometer-based X-ray diffraction data-collection at room temperature and under cryogenic conditions for soluble and membrane-protein crystals grown in or transferred to these plates. We validated the Mylar in situ method using crystals of the water-soluble proteins hen egg-white lysozyme and sperm whale myoglobin as well as the 7-transmembrane protein bacteriorhodopsin from Haloquadratum walsbyi. In conjunction with current developments at synchrotrons, this approach promises high-resolution structural studies of membrane proteins to become faster and more routine. We introduce the Mylar in situ method that uses Mylar-based sandwich plates, which are advantageous over established designs. Cognate holders make the method robust and versatile and allow for automated crystal imaging, screening, and goniometer-based X-ray diffraction data collection at room temperature and under cryogenic conditions for soluble and membrane-protein crystals grown in or transferred to these plates. |
| Databáze: | OpenAIRE |
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