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Title: Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

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. Furthermore, 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:
 [1] ;  [1] ;  [2] ;  [3] ;  [4] ;  [1] ;  [5] ;  [6] ;  [1] ;  [2] ;  [2] ;  [3] ;  [2] ;  [7] ;  [7] ;  [7] ;  [3] ;  [1] ;  [1] ;  [5] more »;  [2] ;  [1] ;  [5] ;  [5] ;  [5] ;  [7] ;  [8] ;  [9] ;  [10] ;  [2] ;  [11] ;  [1] ;  [5] ;  [3] ;  [3] ;  [2] ;  [2] ;  [7] ;  [12] ;  [1] « less
  1. Paul Scherrer Inst. (PSI), Villigen (Switzerland)
  2. Arizona State Univ., Tempe, AZ (United States)
  3. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  4. RIKEN SPring-8 Center, Hyogo (Japan)
  5. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  6. RIKEN SPring-8 Center, Hyogo (Japan); PRESTO, JST, Saitama (Japan)
  7. Univ. of Gothenburg, Gothenburg (Sweden)
  8. RIKEN/SPring-8 Center, Hyogo (Japan)
  9. RIKEN/SPring-8 Center, Hyogo (Japan); Kyoto Univ., Kyoto (Japan)
  10. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
  11. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
  12. Paul Scherrer Inst. (PSI), Villigen (Switzerland); ETH Zurich, Zurich (Switzerland)
Publication Date:
OSTI Identifier:
1313008
Grant/Contract Number:
AC02-76SF00515
Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Research Org:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES