Observation of a single protein by ultrafast X-ray diffraction

Tomas Ekeberg; Dameli Assalauova; Johan Bielecki; Rebecca Boll; Benedikt J. Daurer; Lutz A. Eichacker; Linda E. Franken; Davide E. Galli; Luca Gelisio; Lars Gumprecht; Laura H. Gunn; Janos Hajdu; Robert Hartmann; Dirk Hasse; Alexandr Ignatenko; Jayanath Koliyadu; Olena Kulyk; Ruslan Kurta; Markus Kuster; Wolfgang Lugmayr; Jannik Lübke; Adrian P. Mancuso; Tommaso Mazza; Carl Nettelblad; Yevheniy Ovcharenko; Daniel E. Rivas; Max Rose; Amit K. Samanta; Philipp Schmidt; Egor Sobolev; Nicusor Timneanu; Sergey Usenko; Daniel Westphal; Tamme Wollweber; Lena Worbs; Paul Lourdu Xavier; Hazem Yousef; Kartik Ayyer; Henry N. Chapman; Jonas A. Sellberg; Carolin Seuring; Ivan A. Vartanyants; Jochen Küpper; Michael Meyer; Filipe R. N. C. Maia

doi:10.1038/s41377-023-01352-7

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Observation of a single protein by ultrafast X-ray diffraction

Article

- Observation of a single protein by ultrafast X-ray diffraction
- Light: Science & Applications Vol. 13, Issue 1, Pages: 80-90(2024)
- Affiliations：
- Author bio：
  
  ayfilipe.maia@icm.uu.se
- Funds：
- DOI：10.1038/s41377-023-01352-7
  CLC：
- Received：03 November 2023，
  
  Revised：05 December 2023，
  
  Accepted：2023-12-06，
  
  Published Online：12 January 2024，
  
  Published：2024-12
- Accepted：
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Tomas Ekeberg, Dameli Assalauova, Johan Bielecki, et al. Observation of a single protein by ultrafast X-ray diffraction[J]. Light: science & applications, 2024, 13(1): 80-90.
DOI：

Tomas Ekeberg, Dameli Assalauova, Johan Bielecki, et al. Observation of a single protein by ultrafast X-ray diffraction[J]. Light: science & applications, 2024, 13(1): 80-90. DOI： 10.1038/s41377-023-01352-7.

摘要

Abstract

The idea of using ultrashort X-ray pulses to obtain images of single proteins frozen in time has fascinated and inspired many. It was one of the arguments for building X-ray free-electron lasers. According to theory

the extremely intense pulses provide sufficient signal to dispense with using crystals as an amplifier

and the ultrashort pulse duration permits capturing the diffraction data before the sample inevitably explodes. This was first demonstrated on biological samples a decade ago on the giant mimivirus. Since then

a large collaboration has been pushing the limit of the smallest sample that can be imaged. The ability to capture snapshots on the timescale of atomic vibrations

while keeping the sample at room temperature

may allow probing the entire conformational phase space of macromolecules. Here we show the first observation of an X-ray diffraction pattern from a single protein

that of

Escherichia coli

GroEL which at 14 nm in diameter is the smallest biological sample ever imaged by X-rays

and demonstrate that the concept of diffraction before destruction extends to single proteins. From the pattern

it is possible to determine the approximate orientation of the protein. Our experiment demonstrates the feasibility of ultrafast imaging of single proteins

opening the way to single-molecule time-resolved studies on the femtosecond timescale.

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references

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