Room-temperature macromolecular serial crystallography using synchrotron radiation

Francesco Stellato, Dominik Oberthür, Mengning Liang, Richard Bean, Cornelius Gati, Oleksandr Yefanov, Anton Barty, Anja Burkhardt, Pontus Fischer, Lorenzo Galli, Richard Kirian, Jan Meyer, Saravanan Panneerselvam, Chun Hong Yoon, Fedor Chervinskii, Emily Speller, Thomas A. White, Christian Betzel, Alke Meents, Henry N. Chapman

Research output: Contribution to journalArticle

118 Citations (Scopus)

Abstract

A new approach for collecting data from many hundreds of thousands of microcrystals using X-ray pulses from a free-electron laser has recently been developed. Referred to as serial crystallography, diffraction patterns are recorded at a constant rate as a suspension of protein crystals flows across the path of an X-ray beam. Events that by chance contain single-crystal diffraction patterns are retained, then indexed and merged to form a three-dimensional set of reflection intensities for structure determination. This approach relies upon several innovations: an intense X-ray beam; a fast detector system; a means to rapidly flow a suspension of crystals across the X-ray beam; and the computational infrastructure to process the large volume of data. Originally conceived for radiation-damage-free measurements with ultrafast X-ray pulses, the same methods can be employed with synchrotron radiation. As in powder diffraction, the averaging of thousands of observations per Bragg peak may improve the ratio of signal to noise of low-dose exposures. Here, it is shown that this paradigm can be implemented for room-temperature data collection using synchrotron radiation and exposure times of less than 3ms. Using lysozyme microcrystals as a model system, over 40 000 single-crystal diffraction patterns were obtained and merged to produce a structural model that could be refined to 2.1Å resolution. The resulting electron density is in excellent agreement with that obtained using standard X-ray data collection techniques. With further improvements the method is well suited for even shorter exposures at future and upgraded synchrotron radiation facilities that may deliver beams with 1000 times higher brightness than they currently produce.

Original languageEnglish (US)
Pages (from-to)204-212
Number of pages9
JournalIUCrJ
Volume1
DOIs
StatePublished - Jun 24 2014
Externally publishedYes

Fingerprint

Crystallography
Synchrotrons
Synchrotron radiation
crystallography
synchrotron radiation
X-Rays
Radiation
X rays
Temperature
room temperature
Diffraction patterns
Microcrystals
x rays
diffraction patterns
microcrystals
Suspensions
Single crystals
Electrons
Powder Diffraction
Crystals

Keywords

  • CrystFEL
  • microfocus beamline
  • radiation damage
  • room-temperature protein crystallography
  • serial crystallography

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)
  • Condensed Matter Physics
  • Biochemistry

Cite this

Stellato, F., Oberthür, D., Liang, M., Bean, R., Gati, C., Yefanov, O., ... Chapman, H. N. (2014). Room-temperature macromolecular serial crystallography using synchrotron radiation. IUCrJ, 1, 204-212. https://doi.org/10.1107/S2052252514010070

Room-temperature macromolecular serial crystallography using synchrotron radiation. / Stellato, Francesco; Oberthür, Dominik; Liang, Mengning; Bean, Richard; Gati, Cornelius; Yefanov, Oleksandr; Barty, Anton; Burkhardt, Anja; Fischer, Pontus; Galli, Lorenzo; Kirian, Richard; Meyer, Jan; Panneerselvam, Saravanan; Yoon, Chun Hong; Chervinskii, Fedor; Speller, Emily; White, Thomas A.; Betzel, Christian; Meents, Alke; Chapman, Henry N.

In: IUCrJ, Vol. 1, 24.06.2014, p. 204-212.

Research output: Contribution to journalArticle

Stellato, F, Oberthür, D, Liang, M, Bean, R, Gati, C, Yefanov, O, Barty, A, Burkhardt, A, Fischer, P, Galli, L, Kirian, R, Meyer, J, Panneerselvam, S, Yoon, CH, Chervinskii, F, Speller, E, White, TA, Betzel, C, Meents, A & Chapman, HN 2014, 'Room-temperature macromolecular serial crystallography using synchrotron radiation', IUCrJ, vol. 1, pp. 204-212. https://doi.org/10.1107/S2052252514010070
Stellato F, Oberthür D, Liang M, Bean R, Gati C, Yefanov O et al. Room-temperature macromolecular serial crystallography using synchrotron radiation. IUCrJ. 2014 Jun 24;1:204-212. https://doi.org/10.1107/S2052252514010070
Stellato, Francesco ; Oberthür, Dominik ; Liang, Mengning ; Bean, Richard ; Gati, Cornelius ; Yefanov, Oleksandr ; Barty, Anton ; Burkhardt, Anja ; Fischer, Pontus ; Galli, Lorenzo ; Kirian, Richard ; Meyer, Jan ; Panneerselvam, Saravanan ; Yoon, Chun Hong ; Chervinskii, Fedor ; Speller, Emily ; White, Thomas A. ; Betzel, Christian ; Meents, Alke ; Chapman, Henry N. / Room-temperature macromolecular serial crystallography using synchrotron radiation. In: IUCrJ. 2014 ; Vol. 1. pp. 204-212.
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