Abstract
Free electron lasers (FELs) provide X-ray pulses in the femtosecond time domain with up to 1012 higher photon flux than synchrotrons and open new avenues for the determination of difficult to crystallize proteins, like large complexes and human membrane proteins. While the X-ray pulses are so strong that they destroy any solid material, the crystals diffract before they are destroyed. The most successful application of FELs for biology has been the method of serial femtosecond crystallography (SFX) where nano or microcrystals are delivered to the FEL beam in a stream of their mother liquid at room temperature, which ensures the replenishment of the sample before the next X-ray pulse arrives. New injector technology allows also for the delivery of crystal in lipidic cubic phases or agarose, which reduces the sample amounts for an SFX data set by two orders of magnitude. Time-resolved SFX also allows for analysis of the dynamics of biomolecules, the proof of principle being recently shown for light-induced reactions in photosystem II and photoactive yellow protein. An SFX data sets consist of thousands of single crystal snapshots in random orientations, which can be analyzed now "on the fly" by data analysis programs specifically developed for SFX, but de-novo phasing is still a challenge, that might be overcome by two-color experiments or phasing by shape transforms.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 255-272 |
| Number of pages | 18 |
| Journal | Protein and Peptide Letters |
| Volume | 23 |
| Issue number | 3 |
| State | Published - Mar 1 2016 |
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Keywords
- Femtosecond crystallography
- Free electron lasers
- GPCRs
- Membrane proteins
- Photosystem I
- Photosystem II
ASJC Scopus subject areas
- Biochemistry
- Structural Biology
Cite this
Serial femtosecond crystallography opens new avenues for structural biology. / Coe, Jesse; Fromme, Petra.
In: Protein and Peptide Letters, Vol. 23, No. 3, 01.03.2016, p. 255-272.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Serial femtosecond crystallography opens new avenues for structural biology
AU - Coe, Jesse
AU - Fromme, Petra
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Free electron lasers (FELs) provide X-ray pulses in the femtosecond time domain with up to 1012 higher photon flux than synchrotrons and open new avenues for the determination of difficult to crystallize proteins, like large complexes and human membrane proteins. While the X-ray pulses are so strong that they destroy any solid material, the crystals diffract before they are destroyed. The most successful application of FELs for biology has been the method of serial femtosecond crystallography (SFX) where nano or microcrystals are delivered to the FEL beam in a stream of their mother liquid at room temperature, which ensures the replenishment of the sample before the next X-ray pulse arrives. New injector technology allows also for the delivery of crystal in lipidic cubic phases or agarose, which reduces the sample amounts for an SFX data set by two orders of magnitude. Time-resolved SFX also allows for analysis of the dynamics of biomolecules, the proof of principle being recently shown for light-induced reactions in photosystem II and photoactive yellow protein. An SFX data sets consist of thousands of single crystal snapshots in random orientations, which can be analyzed now "on the fly" by data analysis programs specifically developed for SFX, but de-novo phasing is still a challenge, that might be overcome by two-color experiments or phasing by shape transforms.
AB - Free electron lasers (FELs) provide X-ray pulses in the femtosecond time domain with up to 1012 higher photon flux than synchrotrons and open new avenues for the determination of difficult to crystallize proteins, like large complexes and human membrane proteins. While the X-ray pulses are so strong that they destroy any solid material, the crystals diffract before they are destroyed. The most successful application of FELs for biology has been the method of serial femtosecond crystallography (SFX) where nano or microcrystals are delivered to the FEL beam in a stream of their mother liquid at room temperature, which ensures the replenishment of the sample before the next X-ray pulse arrives. New injector technology allows also for the delivery of crystal in lipidic cubic phases or agarose, which reduces the sample amounts for an SFX data set by two orders of magnitude. Time-resolved SFX also allows for analysis of the dynamics of biomolecules, the proof of principle being recently shown for light-induced reactions in photosystem II and photoactive yellow protein. An SFX data sets consist of thousands of single crystal snapshots in random orientations, which can be analyzed now "on the fly" by data analysis programs specifically developed for SFX, but de-novo phasing is still a challenge, that might be overcome by two-color experiments or phasing by shape transforms.
KW - Femtosecond crystallography
KW - Free electron lasers
KW - GPCRs
KW - Membrane proteins
KW - Photosystem I
KW - Photosystem II
UR - http://www.scopus.com/inward/record.url?scp=84960078639&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84960078639&partnerID=8YFLogxK
M3 - Article
C2 - 26786767
AN - SCOPUS:84960078639
VL - 23
SP - 255
EP - 272
JO - Protein and Peptide Letters
JF - Protein and Peptide Letters
SN - 0929-8665
IS - 3
ER -