Abstract
Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy 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.
Original language | English |
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Pages (from-to) | 421-430 |
Number of pages | 10 |
Journal | IUCrJ |
Volume | 2 |
DOIs | |
Publication status | Published - Jul 1 2015 |
Keywords
- coherent X-ray diffractive imaging
- femtosecond studies
- free-electron laser
- membrane proteins
- nanocrystals
- protein complexes
- serial femtosecond crystallography
- viscous crystal delivery
ASJC Scopus subject areas
- Chemistry(all)
- Biochemistry
- Materials Science(all)
- Condensed Matter Physics