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 languageEnglish (US)
Pages (from-to)255-272
Number of pages18
JournalProtein and Peptide Letters
Volume23
Issue number3
StatePublished - Mar 1 2016

Fingerprint

Crystallography
Free electron lasers
Lasers
X-Rays
Electrons
X rays
Crystals
Microcrystals
Synchrotrons
Photosystem II Protein Complex
Biomolecules
Photons
Diptera
Crystal orientation
Sepharose
Laser beams
Laser pulses
Membrane Proteins
Proteins
Color

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 journalArticle

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