Description
Serial femtosecond crystallography (SFX) has opened a new era in crystallography by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 µg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.
Details
Title
- A novel inert crystal delivery medium for serial femtosecond crystallography
Contributors
- Conrad, Chelsie (Author)
- Basu, Shibom (Author)
- James, Daniel (Author)
- Wang, Dingjie (Author)
- Schaffer, Alexander (Author)
- Roy Chowdhury, Shatabdi (Author)
- Zatsepin, Nadia (Author)
- Aquila, Andrew (Author)
- Coe, Jesse (Author)
- Gati, Cornelius (Author)
- Hunter, Mark S. (Author)
- Koglin, Jason E. (Author)
- Kupitz, Christopher (Author)
- Nelson, Garrett (Author)
- Subramanian, Ganesh (Author)
- White, Thomas A. (Author)
- Zhao, Yun (Author)
- Zook, James (Author)
- Boutet, Sebastien (Author)
- Cherezov, Vadim (Author)
- Spence, John (Author)
- Fromme, Raimund (Author)
- Weierstall, Uwe (Author)
- Fromme, Petra (Author)
- Department of Chemistry and Biochemistry (Contributor)
- Biodesign Institute (Contributor)
- Applied Structural Discovery (Contributor)
- College of Liberal Arts and Sciences (Contributor)
- Department of Physics (Contributor)
- School of Molecular Sciences (Contributor)
Date Created
The date the item was original created (prior to any relationship with the ASU Digital Repositories.)
2015-06-30
Resource Type
Collections this item is in
Identifier
- Digital object identifier: 10.1107/S2052252515009811
- Identifier Value2052-2525
Note
- View the article as published at http://journals.iucr.org/m/issues/2015/04/00/mf5010/index.html
Citation and reuse
Cite this item
This is a suggested citation. Consult the appropriate style guide for specific citation guidelines.
Conrad, C. E., Basu, S., James, D., Wang, D., Schaffer, A., Roy-Chowdhury, S., . . . Fromme, P. (2015). A novel inert crystal delivery medium for serial femtosecond crystallography. IUCrJ, 2(4), 421-430. doi:10.1107/s2052252515009811