The DLAB JRA (JRA7) exists to develop, produce and evaluate systems and protocols for deuterium labelling of biological macromolecules with the aim of enhancing the scope and interdisciplinarity of biological neutron scattering at large scale facilities throughout Europe. This initiative has focused on a number of key issues that have, during the period of the JRA, had a decisive impact on the application of neutron methods for the study of biological systems. Central to this has been the exploitation, throughout Europe, of the capabilities of the ILL-EMBL Deuteration Laboratory in Grenoble to enable more efficient production of selectively and non-selectively H/D-labelled cells, proteins, nucleic acids and other biomolecules. A measure of the importance and impact of this area has been demonstrated (i) by a very clear increase in the requests for facility beam time for studies involving deuterated macromolecules (ii) by the fact that the concept of a Deuteration Laboratory has been duplicated widely at neutron scattering facilities throughout the world (iii) through the results that are now coming into press that would not have been possible without the developments that have occurred as a result of this JRA. A further key element has been training of young scientists in this area – here the impact of this whole initiative is again demonstrable at an international level.

Results

• Improved cost effectiveness of biological isotope labelling

• Development of methods for selectively deuterating specific amino acids in proteins

• Development of methods for deuteration of specific nucleotides in nucleic acids

• Development of methods for reverse labelling: hydrogen labelling in deuterated protein

• Development of novel organisms for deuteration

Applications

JRA7’s scientific tasks have all been successfully completed , achieving major progress with key methodologies for macromolecular deuteration. In general terms, these include the development of methods that have improved the cost effectiveness of biological isotope labelling, as well as methods that have introduced new capabilities for biological neutron scattering. Examples include the ability to selectively deuterate specific amino acids in otherwise hydrogenated protein, methods for selec tive labelling of nucleic acids, as well as methods for hydrogen labelling in deuterated protein (reverse labelling – of particular significance for studies of dynamics), labelling of intact organisms, and the use of new organisms in the production of labelled biomolecules. The impact of these developments is now being felt in the user communities, with increasing numbers of neutron beamtime applications that are making use of these methods. The output in the scientific literature is becoming increasingly marked.

Involvement of partners

All of the partners involved in JRA7 have provided complementary expertise. At the ILL, staff have provided key expertise in molecular biology, adaptation, purification, and characterisation using advanced platforms (eg mass spectroscopy) . At the EMBL, expertise in proteomics has been exploited for an understanding and optimisation of growth conditions for bacteria that are used to produce protein. The CEA has brought in unique expertise on the use of hydrogen incoherent scattering for the study of dynamics, and MPI (Martinsried) has brought expertise on the use of novel organisms for labelling.

Meetings

In addition to the normal meetings of the partners (either through scheduled events at partner institutes, or regular email and telephone correspondence), JRA7 activities have been central to a number of other meetings. Examples of meetings where JRA7-related activity has been presented include:

• HERCULES Specialised course on Advances and new applications for structural biology, ESRF Grenoble (8th Oct 2007)
• Neutrons in Biology, Rutherford-Appleton Laboratory, Oxfordshire (11-13th July 2007) (NMI3 supported). (Special issue of European Biophysics Journal).
• Proteins in action: neutron scattering to investigate biomolecules at physiological conditions, Perugia (6-8th June 2007). (NMI3 supported). (Special issue of European Biophysics Journal).
• Soft Condensed Matter workshop, Institut Laue Langevin, Grenoble, France, 22-24 November 2006.
• XX Congress of the International Union of Crystallography, Florence.
• Bioforum meeting, Coseners House, Didcot, UK (6-7 April 2006).
• International Conference on Neutron Scattering, Sydney, Australia (27th Nov -2nd Dec. 2005)
• Neutrons in biology, a satellite meeting to IUPAB/EBSA Conference, Montpellier (27th-31st Aug 2005), Grenoble, France (August/September 2005) (NMI3 supported). (Special issue of European Biophysics Journal).