Oral 23rd International Society of Magnetic Resonance Conference 2023

Enabling biomolecular investigations using NMR spectroscopy: isotopic labelling strategies of the Australian National Deuteration Facility. (#37)

Karyn L Wilde 1 , Anthony P Duff 1 , Rao R Yepuri 1 , Tamim Darwish 1
  1. National Deuteration Facility - ANSTO, Lucas Heights, NSW, Australia

Molecular deuteration of organic compounds and biomolecules is an essential prerequisite in many 2H (deuterium) NMR, infrared, mass spectroscopy, and neutron scattering studies. To reduce spectral complexity and signal linewidths in both solution and solid-state NMR spectra, the application of various stable isotope labelling schemes for proteins has been well established within the biomolecular NMR community. The National Deuteration Facility (NDF) at ANSTO is the only facility of its kind in the Southern Hemisphere, with the specialised expertise and infrastructure to produce a broad range of deuterated molecules through both biological and chemical techniques that may otherwise not be available commercially. The NDF enables biomolecular investigations using NMR through the operation of a proposal-based user program (offering different modes of access) for provision of bespoke deuterated and multiple-isotope labelled molecules to researchers and industry.

The NDF utilises their reliable and robust methods for the deuteration of a broad range of proteins by high-cell density, high-yield recombinant expression in Escherichia coli BL21 using a simple defined method [1]. These methods have been adapted to produce uniformly double (13C/15N, 2H/15N) and triple labelled (2H/13C/15N) proteins, as well as 13C- Ala, Ile, Leu Val (AILV) methyl labelled proteins and proteins with selective amino-acid labelling, for the support of structural and functional investigations. Selected examples of protein labelling and their application will be highlighted [2-8].

Molecules produced through chemical deuteration and used in neutron studies such as ionic liquids, unsaturated lipids, uncommon surfactants, sugars, and heterocyclic and aromatic compounds can also be applied to NMR studies, enabling different and complementary experimental approaches for your research. The deuteration of novel molecules by the NDF with NMR applications will be described, including but not limited to applications for studying self-assembled gels and digestion of medium chain triglycerides [9-10].

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