Department of Biochemistry University of Oxford Department of Biochemistry
University of Oxford
South Parks Road
Oxford OX1 3QU

Tel: +44 (0)1865 613200
Fax: +44 (0)1865 613201
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Elspeth Garman
Radiation damage in X-ray crystallography and elemental analysis of proteins

Co-workers: Mr. Markus Gerstel, Miss. Katharina Jungnickel, Mr. Jonathan Brooks-Bartlett, Mr. Oliver Starkey

Our research focuses on developing improved methods for macromolecular crystallography (MX) to enable problems not previously accessible to structure solution to be tackled. This work currently includes studies on 100K and room temperature (RT) radiation damage and optimising strategies for in-house Sulphur and Phosphorus-SAD phase determinations.

Radiation damage (Fig 1) to the sample is an inherent problem when utilising ionising X-radiation in MX, even during cryocrystallographic (100K) data collections. We seek to understand the physical and chemical basis of the damage, and to find mitigation strategies in order to optimise our data collections and thus maximise the biological information obtained

Following our measurement of 30 MGy for the experimental dose limit for crystals held at 100K, we have investigated the RT dose limit. Surprisingly, the dose tolerated by a RT crystal is dependent on the dose rate according to a positive linear relationship. The addition of scavengers changes the RT intensity loss dependence from first to zeroth order (Fig 2) in dose.

Our method for accurately determining the concentration of trace elements bound in liquid or crystalline protein samples by proton induced X-ray emission (PIXE) is finding ongoing applications (Fig 3), and is being extended to measurements on whole cells.

Publications

  1. Predicting the X-ray lifetime of protein crystals
    Oliver B. Zeldin, Sandor Brockhauser, John Bremridge, James Holton and Elspeth Garman.  PNAS 110, 20551-6, On-line 2/12/13 2013
  2. Structure of arylamine N-acetyltransferase from M. tuberculosis determined by cross-seeding with homologous protein from M. marinum: Triumph over Adversity.
    Areej Abuhammad, Edward D. Lowe, Michael A. McDonough, Patrick D. Shaw Stewart, Stefan A. Kolek, Edith Sim, Elspeth F. Garman. Acta Cryst. D (2013) 69, 1433-1446
  3. RADDOSE-3D: time- and space-resolved modeling of dose in macromolecular crystallography
    Oliver B. Zeldin, Markus Gerstel and Elspeth Garman Journal of Applied Crystallography (2013) 46, 1225-1230
  4. Experimental determination of the radiation dose limit for cryocooled protein crystals. Robin Leslie Owen, Enrique Rudiño-Piñera, Elspeth F. Garman (2006) Proc. Nat. Acad. Sci (2006) 103, 4912-4917
  5. Elemental analysis of proteins by microPIXE. Elspeth Garman and Geoff Grime (2005) Progress in Biophysics and Molecular Biology 89/2, 173-205
More Publications...

Research Images



Figure 1: Radiation damage to an N9 neuraminidase from avian influenza, following 100K data collection at the ESRF from 3 locations on the crystal.
 


Figure 2: Room temperature change in form of the intensity loss dependence with dose on addition of the scavenger 1,4 benzoquinone to crystals of hen egg white lysozyme. Addition of the scavenger increases the dose tolerance by a factor of 9
 

Figure 3: Elemental areal concentration microPIXE maps (500µm x 500µm) obtained by scanning a 1 µm diameter proton beam in x and y across a holoferritin crystal showing: sulphur (left), iron (centre) and cadmium (right) distributions. The iron is localised in the protein crystal, while due to the presence of ammonium sulphate in the solvent, the sulphur is more spread out.
 


Contact: elspeth.garman@bioch.ox.ac.uk
Graduate Student and Postdoctoral Positions: Enquiries with CV welcome
Website: www.bioch.ox.ac.uk/garmangroup