Poster Presentation 2014 International Biophysics Congress

Molecular deuteration enabling structural studies of proteins and biomembranes using neutron scattering and NMR (#483)

Peter J Holden 1 , Tamim A Darwish 1 , Anthony P Duff 1 , Marie Gillon 1 , Anwen Krause-Heuer 1 , Agata Rekas 1 , Karyn L Wilde 1 , Rao Yepuri 1
  1. ANSTO, Kirrawee DC, NSW, Australia

Molecular deuteration significantly increases the options available for structure-function investigations of proteins and biomembranes using neutron scattering and NMR. The National Deuteration Facility has developed high yield (50-100 mg/L) protocols for partial and perdeuteration of proteins for small angle neutron scattering (SANS), neutron reflectometry, crystallography and spectrometry (dynamics). Case studies on the Munc18:Syntaxin neuronal protein complex involved in signal propagation at synapses (1) and on the interaction of Calmodulin with HIV-1 matrix protein (2) will demonstrate the efficacy of contrast variation SANS combined with deuteration in resolving shape and disposition of proteins in complexes. These protocols have also been successfully adapted to achieve high yields of 2H/15N/13C labelled proteins for NMR studies, e.g. of the fungal hydrophobin functional amyloid (3), and extension to include labelling of specific amino acids with 15N/13C against deuteration of all remaining amino acids is under-way. Likewise, deuteration combined with neutron reflectometry has enabled characterisation of the surfactant behaviour of short peptides at the air-water interface (4). These capabilities are complemented by our Chemical Deuteration activities which provide a range of phospholipids (head or tail deuterated DOPC, DPPC, DMPC, POPC and the archaeal diphytanoyl- glycerol PC (5)) and detergents (n-dodecyl--D-maltopyranoside and n-octyl--D-glucoside) relevant, respectively, to biomembrane bilayer and biosensor investigations, and to membrane protein (and drug delivery) SANS studies using nanodiscs or liposomes. An overview using applications of such molecules deuterated at the NDF will be used to communicate the potential of deuteration strategies to facilitate complex structural studies.






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  5. N.R. Yepuri et al., Chem Phys. Lipids In Press (2014).