Room-temperature macromolecular serial crystallography using synchrotron radiation
| Autor: | Saravanan Panneerselvam, Alke Meents, Pontus Fischer, Henry N. Chapman, Richard A. Kirian, Anja Burkhardt, Mengning Liang, Oleksandr Yefanov, Chun Hong Yoon, Lorenzo Galli, Fedor Chervinskii, Christian Betzel, Francesco Stellato, Jan Meyer, Dominik Oberthür, Emily M. Speller, Cornelius Gati, Anton Barty, Richard Bean, Thomas A. White |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Diffraction
CrystFEL Computer science Synchrotron radiation Bragg peak Biochemistry law.invention law ddc:530 General Materials Science serial crystallography lcsh:Science Detector Resolution (electron density) Settore FIS/07 General Chemistry Condensed Matter Physics Laser Research Letters Crystallography radiation damage lcsh:Q room-temperature protein crystallography microfocus beamline Ultrashort pulse Powder diffraction |
| Zdroj: | IUCrJ, Vol 1, Iss 4, Pp 204-212 (2014) IUCrJ IUCrJ 1(4), 204-212 (2014). doi:10.1107/S2052252514010070 |
| ISSN: | 2052-2525 |
| Popis: | The room-temperature structure of lysozyme is determined using 40000 individual diffraction patterns from micro-crystals flowing in liquid suspension across a synchrotron microfocus beamline. A new approach for collecting data from many hundreds of thousands of microcrystals using X-ray pulses from a free-electron laser has recently been developed. Referred to as serial crystallography, diffraction patterns are recorded at a constant rate as a suspension of protein crystals flows across the path of an X-ray beam. Events that by chance contain single-crystal diffraction patterns are retained, then indexed and merged to form a three-dimensional set of reflection intensities for structure determination. This approach relies upon several innovations: an intense X-ray beam; a fast detector system; a means to rapidly flow a suspension of crystals across the X-ray beam; and the computational infrastructure to process the large volume of data. Originally conceived for radiation-damage-free measurements with ultrafast X-ray pulses, the same methods can be employed with synchrotron radiation. As in powder diffraction, the averaging of thousands of observations per Bragg peak may improve the ratio of signal to noise of low-dose exposures. Here, it is shown that this paradigm can be implemented for room-temperature data collection using synchrotron radiation and exposure times of less than 3 ms. Using lysozyme microcrystals as a model system, over 40 000 single-crystal diffraction patterns were obtained and merged to produce a structural model that could be refined to 2.1 Å resolution. The resulting electron density is in excellent agreement with that obtained using standard X-ray data collection techniques. With further improvements the method is well suited for even shorter exposures at future and upgraded synchrotron radiation facilities that may deliver beams with 1000 times higher brightness than they currently produce. |
| Databáze: | OpenAIRE |
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