Protein Crystallography: Xenon Derivative

Rob Kehoe & James Nicholson

We are developing facilities allowing the use of xenon derivatives (at room temp. and 100K) for MIR and Anomalous Dispersion experiments on the protein crystallography stations at the SRS, Daresbury.


  • Derivative protein crystals can be produced by pressurising native crystals with xenon gas. Modification of the mother liquor to determine soaking conditions is avoided.

  • The number of binding sites, and their occupancies can be changed by altering the gas pressure, thus, several derivatives could be produced from the same crystal. These sites often differ from metal binding sites.

  • Xenon atoms interact weakly with protein; isomorphism of the derivative with the native is high.

  • Xenon binding is often reversible; the same crystal could be used for a further heavy atom soak.


  • Systematic errors in measured intensities due to absorption by the crystal, mother liquor and pressurising gas.
  • Accelerated radiation damage to unfrozen crystals.
  • Safety implications associated with high pressures.
  • Possible formation of hydrates at low temperatures and high pressures.


    XENON (Xe)
    *Atomic no54*
    *Melting point-112*
    *Boiling point-108*
    *Relative density (Gas)4.5*
    *Relative density (liquid)1.5*
    *Molecular weight131*
    *Solubility in water644 mg/l*
    Absorption edges (Å and (kev))
    (34.5614)(5.4528)(5.1037)(4.7822) (1.1446)

    Text file of anomalous scattering coefficients f' and f'' as a Function of energy


    Schoenborn et al determine a xenon binding site in sperm whale metmyoglobin by collecting x-ray diffraction data to a resolution of 2.8Å from crystals pressurised to 2.5 atmospheres.

    Tilton et al varied the number of bound xenon atoms by altering the sample pressure.

    Vitali et al demonstrate the use of xenon as a heavy atom, for the first time, in determining phases for the structure of sperm whale metmyoglobin, with the technique of isomorphous replacement with anomalous scattering (SIRAS).

    Schiltz et al use xenon as an alternative to heavy metal derivatives; for the first time calculate phases of a protein of unknown structure using a xenon derivative.

    Soltis et al for the first time, demonstrate a method for preparing cry-cooled xenon derivatised protein crystals.


    We are developing a xenon cell at Daresbury Laboratory for collectiong room temp. data from xenon derivative crystals in the 0 to 25 bar pressure range. It is a very simple but effective device based on a commercially available compression fitting. The Room Temperature Cell Mounted on a Goniometer Head.

    Schematic Diagram of the Xenon Cell.

    The Various Components of the Room Temperature Xenon Cell.

    The Xenon Cell Mounted on the Goniostat of
    Station 7.2 at the SRS.

    Users of the xenon pressure cell at Daresbury are required to complete a class 3 risk assessment, including a description of the hazard and safe system of work, prior to their beamtime.

    Future Developments


  • Ethan A Merritt 1996-1999/ / Biomolecular Structure Center at UW


  • Tutorial by Mike Soltis of the SSRL
  • Xenon and Krypton at LURE
  • Xenon Pressure Cells at the SSRL


  • Binding of Xenon to Sperm Whale Myoglobin. Schoenborn B.P.; Watson, H.C.; Kendrew, J.C. (1965). Nature, 207, 28-30.

  • Cavities in proteins: structure of a metmyoglobin-xenon complex solved to 1.9&Angring.; Tilton, R.F.; Kuntz, L.D.; Pesko, G.A. (1984) Biochemistry 23. 2849-2857.

  • Using Xenon as a Heavy Atom for Determining Phases in Sperm Whale Metmyoglobin. Vitali, J.; Robbins, A.H.; Almo, S.C.; Tilton, R.F. (1991). Journal of Applied Crystallography, 24, 931-935.

  • On the Preparation and X-ray Data Collection of Isomorphous Xenon Derivatives. Schiltz, M.; Prange, T.; Fourme, R. (1994). Journal of Applied Crystallography, 27, 950-960.

  • Successful flash-cooling of xenon-derivatised myoglobin crystals. Soltis, S.M.; Stowell, M.H.B.; Wiener, M.C.; Philips, G.N.; Rees, D.C. (1997).Journal of Applied Crystallography, 30, 190-194.

  • Freeze-Trapping Isomorphous Xenon Derivatives of Protein Crystals. Sauer, O.; Schmidt, A.; Kratky, C. (1997). Journal of Applied Crystallography, 30, 476-486.

  • Protein Crystallography at Ultra-Short Wavelengths: Feasibility Study of Anomalous Dispersion Experiments at the Xenon K-Edge. Schiltz, M., Kvick, A., Svensson, O., Shepard, W., De LaFortelle, E., Prange, T., Kahn, R. & Fourme, R. (1997). Journal of Synchrotron Radiation , 4, 287-297.

  • A method to stabilize reduced and/or gas treated protein crystals by flash cooling under a controlled atmospher
    Xavier Vermede J. App. Cryst, (1999) 32(3) 505-509

  • Better structures from better data through better methods: a review of developments in de novo macromolecular phasing techniques and associated instrumentation at LURE. Fourme, R. et al (1999) J. Synch. Rad. 6(4) 834-844


  • Teng et al (1990) J. Appl. Cryst. 23. 387-391


    This page was last updated 12th May. 1999