Journal article
A routine method for cloning, expressing and purifying Aβ(1-42) for structural NMR studies
DK Weber, MA Sani, JD Gehman
Amino Acids | SPRINGER WIEN | Published : 2014
Abstract
Nuclear magnetic resonance (NMR) is a key technology in the biophysicist's toolbox for gaining atomic-level insight into structure and dynamics of biomolecules. Investigation of the amyloid-β peptide (Aβ) of Alzheimer's disease is one area where NMR has proven useful, and holds even more potential. A barrier to realizing this potential, however, is the expense of the isotopically enriched peptide required for most NMR work. Whereas most biomolecular NMR studies employ biosynthetic methods as a very cost-effective means to obtain isotopically enriched biomolecules, this approach has proven less than straightforward for Aβ. Furthermore, the notorious propensity of Aβ to aggregate during purifi..
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Awarded by RMIT University
Funding Acknowledgements
The authors would like to sincerely thank Dr. Nick Williamson, Paul O'Donnell and Michael Leeming for discussions regarding ESI-MS acquisition and analysis, John Karas for advice on HPLC purification, and Professor Anthony Wedd and Dr. Zhiguang Xiao for allowing access to equipment required for cell-culture work. J. Gehman was partially funded by ARC Future Fellowship FT0991558 for this work. Circular Dichroism and Dynamic Light Scattering instruments were funded by a LIEF grant LE120100186 to G. Bryant (RMIT) and J. Gehman. D. Weber is thankful for an Australian Postgraduate Award PhD scholarship and Dowd Foundation Postgraduate Research Scholarship for Neuroscience.