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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

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.
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    Title
    • Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography
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    Date Created
    2016-08-22
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    Identifier
    • Digital object identifier: 10.1038/ncomms12314
    • Identifier Type
      International standard serial number
      Identifier Value
      2041-1723
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    • The final version of this article, as published in Nature Communications, can be viewed online at: https://www.nature.com/articles/ncomms12314

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    Nogly, P., Panneels, V., Nelson, G., Gati, C., Kimura, T., Milne, C., . . . Standfuss, J. (2016). Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography. Nature Communications, 7, 12314. doi:10.1038/ncomms12314

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