Oral Presentation Australian & New Zealand Society of Magnetic Resonance Conference 2017

Using 5D-APSY for complete protein backbone assignment in solid-state (#67)

Henry Orton 1 , Jan Stanek 2 , Guido Pintacuda 2
  1. Australian National Univeristy, Braddon, ACT, Australia
  2. Centre de résonance magnétique nucleaire à trés hauts champs, Lyon, France

Automated projection spectroscopy (APSY) is a sparse sampling technique that uses multiple 2D projections of higher dimensional spectra to acquire a peak list within a reasonable time-period. APSY is most commonly applied in solution-state, however the relatively long coherence lifetimes achieved with ultra-fast magic angle spinning (MAS) in solids permit higher sensitivity during long experiments. Here we present the first application of APSY in solids using a 5D proton-detected hNCOCANH experiment on a microcrystalline sample of GB1 at 111 kHz MAS. When coupled with the reverse hNCACONH experiment and a total experiment time of 12 hours, these experiments show great power for sequential assignment. With just these two experimental peak lists, complete and accurate assignment was achieved by the automated software FLYA within a few minutes. In future, using APSY in solid-state has the potential to save time and greatly simplify the process of protein assignment.

  1. S. Hiller, F. Fiorito, K. Wüthrich, G. Wider (2005) Automated projection spectroscopy (APSY), Proc. Natl. Acad. Sci. 102:10876–10881
  2. E. Schmidt, P. Güntert, (2012) A new algorithm for reliable and general NMR resonance assignment. J. Am. Chem. Soc. 134:12817–12829