5J7A : Bacteriorhodopsin ground state structure obtained with Serial Femtosecond Crystallography

  • Przemyslaw Nogly (Contributor)
  • Valerie Panneels (Contributor)
  • G. Nelson (Contributor)
  • Cornelius Gati (Contributor)
  • Tetsunari Kimura (Contributor)
  • Christopher Milne (Contributor)
  • Despina Milathianaki (Contributor)
  • Minoru Kubo (Contributor)
  • Wenting Wu (Contributor)
  • Chelsie E. Conrad (Contributor)
  • Jesse Coe (Contributor)
  • Richard J. Bean (Contributor)
  • Yun Zhao (Contributor)
  • Petra Båth (Contributor)
  • Robert Dods (Contributor)
  • Rajiv Harimoorthy (Contributor)
  • Kenneth R. Beyerlein (Contributor)
  • Jan Rheinberger (Contributor)
  • Daniel James (Contributor)
  • D. P. Deponte (Contributor)
  • Chufeng Li (Contributor)
  • Leonardo Sala (Contributor)
  • Garth J. Williams (Contributor)
  • Mark S. Hunter (Contributor)
  • J. E. Koglin (Contributor)
  • Peter Berntsen (Contributor)
  • Eriko Nango (Contributor)
  • So Iwata (Contributor)
  • Henry N. Chapman (Contributor)
  • Petra Fromme (Contributor)
  • Matthias Frank (Contributor)
  • R. Abela (Contributor)
  • Sébastien Boutet (Contributor)
  • Anton Barty (Contributor)
  • Thomas A. White (Contributor)
  • Uwe Weierstall (Contributor)
  • John Spence (Contributor)
  • Richard Neutze (Contributor)
  • Gebhard F X Schertler (Contributor)
  • Jörg Standfuss (Contributor)



Experimental Technique/Method:X-RAY DIFFRACTION
Release Date:2016-08-31
Deposition Date:2016-04-06
Revision Date:2018-01-24
Molecular Weight:28341.51
Macromolecule Type:Protein
Residue Count:227
Atom Site Count:1792

Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1 mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.
Date made available2016

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