DNP enhanced MAS solid-state NMR of biological systems in their native context

• Applicant: Professor Dr. Bernd Reif
• Subject Area: Structural Biology
• Term: from 2012 to 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 226561002

• Final Report Year: 2019

Final Report Abstract

We have established that membrane proteins in a native environment can be investigated by DNP MAS solid-state NMR. We observe a significant increase in the spectral resolution from 400 MHz to 800 MHz. At the same time, enhancements decay by one order of magnitude. Development in design of polarizing agents is needed to fully exploit DNP experimental results at high magnetic fields for biological applications. The spectral quality of amino acid selective labeled Mistic samples is in principle sufficient to yield sequential assignments for amino acid stretches in a native lipid preparation without the need for purification and reconstitution of the membrane protein.

Publications

• (2013) Cryogenic solid state NMR studies of fibrils of the Alzheimer’s disease amyloid-β peptide: perspectives for DNP. J. Biomol. NMR 56: 359–363
  Lopez del Amo J-M, Schneider D, Loquet A, Lange A, Reif B
  (See online at https://doi.org/10.1007/s10858-013-9755-5)
• (2013) The Mechanism of Denaturation and the Unfolded State of the α-Helical Membrane-Associated Protein Mistic. J. Am. Chem. Soc. 135: 18884−18891
  Jacso T, Bardiaux B, Broecker J, Fiedler S, Barwinkel T, Mainz A, Fink U, Vargas C, Oschkinat H, Keller S, Reif B
  (See online at https://doi.org/10.1021/ja408644f)