Structure determination of an integral membrane protein at room temperature from crystals in situ
| Autor: | Nien-Jen Hu, James Foadi, So Iwata, Hassanul G. Choudhury, Danny Axford, Gwyndaf Evans, Konstantinos Beis, Yilmaz Alguel |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
In situ
Diffraction Materials science Diffusion Analytical chemistry macromolecular substances 010402 general chemistry 01 natural sciences law.invention 03 medical and health sciences Structural Biology law Radiation damage membrane protein Crystallization 030304 developmental biology 0303 health sciences in situ data collection Resolution (electron density) General Medicine Research Papers Synchrotron 0104 chemical sciences multiple data sets X-ray crystallography biological sciences synchrotron beamline |
| Zdroj: | Acta Crystallographica Section D: Biological Crystallography |
| ISSN: | 1399-0047 0907-4449 |
| Popis: | The X-ray structure determination of an integral membrane protein using synchrotron diffraction data measured in situ at room temperature is demonstrated. The structure determination of an integral membrane protein using synchrotron X-ray diffraction data collected at room temperature directly in vapour-diffusion crystallization plates (in situ) is demonstrated. Exposing the crystals in situ eliminates manual sample handling and, since it is performed at room temperature, removes the complication of cryoprotection and potential structural anomalies induced by sample cryocooling. Essential to the method is the ability to limit radiation damage by recording a small amount of data per sample from many samples and subsequently assembling the resulting data sets using specialized software. The validity of this procedure is established by the structure determination of Haemophilus influenza TehA at 2.3 Å resolution. The method presented offers an effective protocol for the fast and efficient determination of membrane-protein structures at room temperature using third-generation synchrotron beamlines. |
| Databáze: | OpenAIRE |
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