Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography Academic Article uri icon

abstract

  • 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.

authors

  • Nogly, P
  • Panneels, V
  • Nelson, G
  • Gati, C
  • Kimura, T
  • Milne, C
  • Milathianaki, D
  • Kubo, M
  • Wu, W
  • Conrad, C
  • Coe, J
  • Bean, R
  • Zhao, Y
  • Båth, P
  • Dods, R
  • Harimoorthy, R
  • Beyerlein, KR
  • Rheinberger, J
  • James, D
  • DePonte, D
  • Li, C
  • Sala, L
  • Williams, GJ
  • Hunter, MS
  • Koglin, JE
  • Berntsen, P
  • Nango, E
  • Iwata, S
  • Chapman, HN
  • Fromme, P
  • Frank, M
  • Abela, R
  • Boutet, S
  • Barty, A
  • White, TA
  • Weierstall, U
  • Spence, J
  • Neutze, R
  • Schertler, G
  • Standfuss, J

publication date

  • 2016