Robin Owen

It is often difficult to obtain large well diffracting crystals. By developing methods and intrumentation for Microfocus MX we aim to enable structure solution for the most challenging crystals be they small, poorly ordered or invisible when cryo-cooled for data collection. 

Hardware developments resulting in smaller, hotter beams combined with the weakly diffracting nature of macromolecular crystals means that radiation damage is often a limiting factor in macromolecular crystallography. In order to maximise the amount of useful information a crystallographer can extract from a crystal, we aim to userstand the processes of radiation damage in crystals at both 100K and room temperature. Complementary methods can provde a means for this in addition to providing valuable biological information. 

Even if an experiment is carried out optimally, it is often only possible to collect a small amount of data from each crystal. Many 10's, 100's or even 1000's of crystals may be required to obtain a complete dataset. We are developing instrumentation for new modes of sample delivery to make efficient use of crystals at both Diamond and X-ray Free Electron Lasers (XFELs). Despite the obvious differences in synchrotron and XFEL data collection, there are many common areas and our approaches aim to exploit these. 

XFELs provide a means of obtaining high resolution time resolved crystallographic structures: a limitation of conventional MX is that the model obtained provides a static snapshot of the reaction of interest. For a complete understanding of how systems function it is vital to be able to watch and understand chemical reactions and biological processes as they occur. Together with a number of collaborators the methods we are helping to develop will enable high throughput Fixed Target Serial Femtosecond Crystallography (FT-SFX). This is an approach that can be used at both synchrtrons and FELs, opening up the field of SFX for difficult to crystallise targets. 

I work with several groups outside Diamond as part of my research. These include:

Mike Hough (University of Essex). Methods for high throughput serial crystallography.

Arwen Pearson (University of Hamburg). We work together on a number of projects focusing on the use of complementary methods and development of novel approaches for time-resolved crystallography. 

Ray Owens, Jo Nettleship (OPPF-UK); Jim Robinson, Ann Morgan and Darren Tomlinson (University of Leeds). Structural studies of clincally important proteins implicated in rheumatoid arthritis.

Dwayne Miller (University of Toronto & University of Hamburg). Methods for serial femtosecond crystallography.

A list of my publications can be found below. Please contact me if you would like a copy of any of the below

2. Where is crystallography going?

3. Affimer proteins inhibit immune complex binding to FcγRIIIa with high specificity through competitive and allosteric modes of action

4. Time zero determination for FEL pump-probe studies based on ultrafast melting of bismuth

5. Low dose fixed target serial synchrotron crystallography

6. The use of haptic interfaces and web services in crystallography: an application for a `screen to beam' interface

7. Exploiting microbeams for membrane protein structure determination

8. Solid Target combined with spectral mapping: approaching 100% hit rates for serial crystallography using XFELs and synchrotrons

9. Current advances in synchrotron radiation instrumentation for MX experiments

10. Radiation damage and derivatization in macromolecular crystallography: a structure factor's perspective

11. Imperfection and radiation damage in protein crystals studied with coherent radiation

12. A modular and compact portable mini-endstation for high precision, high speed fixed target serial crystallography at FELs and synchrotron sources.

13. Fixed target matrix for femtosecond and in situ serial micro-crystallography

14. Polyhedra structures and the evolution of the insect viruses

15. Status of the crystallography beamlines at Diamond Light Source

16. Time-resolved crystallography using the Hadamard transform.

17. Exploiting fast detectors to enter a new dimension in room temperature crystallography

18. Structural and Mechanistic Insights into the PAPS-Independent Sulfotransfer Catalyzed by Bacterial Aryl Sulfotransferase and the Role of the DsbL/DsbI System in its Folding

19. Adhiron: a new and versatile protein scaffold

20. Human cellular retinaldehyde-binding protein has secondary thermal 9-cis-retinal isomerase activity

21. Structure and mechanism of N33/Tusc3, a redox-active subunit of the human oligosaccharyl transferase complex

22. Insights into the mechanism of X-ray induced disulfide bond cleavage in lysozyme crystals based on EPR, optical absorption, and X-ray diffraction studies

23. Clustering procedures for the optimal slection of data sets from multiple crystals in macromolecular crystallography

24. Outrunning free radicals in room temperature macromolecular crystallography

25. X-ray excited optical luminescence of protein crystals: a new tool for studying radiation damage during diffraction data collection

26. In situ macromolecular crystallography using microbeams

27. A sensor/adaptor mechanism for enterovirus uncoating from structures of EV71

28. Four complete turns of a curved 3(10)-helix at atomic resolution: The crystal structure of the peptaibol trichovirin I-4A in polar environment suggests a transition to alpha-helix for membrane function

29. Macromolecular Microcrystallography

30. Revealing low dose radiation damage using single crystal spectroscopy

31. High-speed crystal detection and characterisation using a fast readout detector.

32. Mammalian cell expression, purification, crystallization and microcrystal data collection of Autotaxin/ENPP2, a secreted mammalian glycoprotein

33. Characterization of a C-C hydrolase from Mycobacterium tuberuclosis involved in cholesterol metabolism

34. How baculovirus polyhedra fit square pegs into round holes to robustly package viruses.

35. Combining X-ray crystallography and single crystal spectroscopy to probe enzyme mechanisms.

36. Absorbed dose calculations for macromolecular crystals: improvements to RADDOSE

37. Determination of X-ray flux using silicon pin diodes

38. Colouring Cryo-Cooled Crystals: on-line microspectrophotometry.

39. A novel on-axis multi-mode spectrometer at X10SA of the Swiss Light Source

40. A structural and biochemical basis for PAPS-independent sulfuryl transfer by aryl sulfotransferase from uropathogenic Escherichia coli.

41. Preparation and structure of the charge-transfer intermediate of the transmembrane redox catalyst DsbB

42. Resonance Raman spectroscopy for In-situ monitoring of radiation damage.

44. Experimental determination of the radiation dose limit for cryocooled protein crystals.

45. Effect of Irradiation-Induced Disorder on the Conductivity and Critical Temperature of the Organic Superconductor k-BEDT-TTF₂CuSCN₂.

46. Cryocooling and radiation damage in macromolecular crystallography

47. Parameters affecting the X-ray dose absorbed by macromolecular crystals.

48. A new method for predetermining the diffraction quality of protein crystals: using SOAP as a selection tool.

49. Temperature characteristics of crystal storage devices in a CP100 dry shipping Dewar.