The National Deuteration Facility (NDF) has developed expertise in biological molecular deuteration for the in vivo deuteration of proteins, where all or a fraction of the hydrogen atoms in an expressed protein are replaced by the stable isotope deuterium (2H).
The NDF capability for the production of multiple-labelled proteins using the stable isotopes 2H, 13C and 15N is of particular interest to the NMR community for both solution and solid‑state NMR applications. As with deuterium (2H), the 13C and/or 15N are introduced into the minimal growth medium utilised for biomass production and protein expression using the NDF methods for high-yield recombinant expression of partial or per-deuterated protein in Escherichia coli [1]. Selective-labelling of desired amino acids with 13C and/or 15N in recombinantly expressed proteins has also been demonstrated, with further capability development currently being undertaken at the NDF for other labelling strategies such as selective-labelling with deuteration and methyl labelling.
A brief overview of NDF labelling methods will be presented and the facilities and expertise available to the neutron beam and NMR user communities discussed, highlighted with examples of NMR and neutron investigations utilising deuterated and multiple-labelled proteins.
Access to the NDF is available through an externally refereed proposal scheme. For further detail and information refer to http://www.ansto.gov.au/ndf .
The National Deuteration Facility is partially funded by the National Collaborative Research Infrastructure Strategy (NCRIS), an initiative of the Australian Federal Government.