Powder diffraction from a continuous microjet of submicrometer protein crystals

D. A. Shapiro, H. N. Chapman, D. DePonte, R. B. Doak, P. Fromme, G. Hembree, M. Hunter, S. Marchesini, K. Schmidt, J. Spence, D. Starodub, U. Weierstall

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

Atomic-resolution structures from small proteins have recently been determined from high-quality powder diffraction patterns using a combination of stereochemical restraints and Rietveld refinement [Von Dreele (2007), J. Appl. Cryst. 40, 133-143; Margiolaki et al. (2007), J. Am. Chem. Soc. 129, 11865-11871]. While powder diffraction data have been obtained from batch samples of small crystal-suspensions, which are exposed to X-rays for long periods of time and undergo significant radiation damage, the proof-of-concept that protein powder diffraction data from nanocrystals of a membrane protein can be obtained using a continuous microjet is shown. This flow-focusing aerojet has been developed to deliver a solution of hydrated protein nanocrystals to an X-ray beam for diffraction analysis. This method requires neither the crushing of larger polycrystalline samples nor any techniques to avoid radiation damage such as cryocooling. Apparatus to record protein powder diffraction in this manner has been commissioned, and in this paper the first powder diffraction patterns from a membrane protein, photosystem I, with crystallite sizes of less than 500 nm are presented. These preliminary patterns show the lowest-order reflections, which agree quantitatively with theoretical calculations of the powder profile. The results also serve to test our aerojet injector system, with future application to femtosecond diffraction in free-electron X-ray laser schemes, and for serial crystallography using a single-file beam of aligned hydrated molecules.

Original languageEnglish
Pages (from-to)593-599
Number of pages7
JournalJournal of Synchrotron Radiation
Volume15
Issue number6
DOIs
Publication statusPublished - Oct 3 2008

Keywords

  • Membrane proteins
  • Nanocrystals
  • Protein structure
  • Radiation damage
  • Serial crystallography

ASJC Scopus subject areas

  • Radiation
  • Nuclear and High Energy Physics
  • Instrumentation

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