1. Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.
- Author
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Andreas LB, Stanek J, Le Marchand T, Bertarello A, Cala-De Paepe D, Lalli D, Krejčíková M, Doyen C, Öster C, Knott B, Wegner S, Engelke F, Felli IC, Pierattelli R, Dixon NE, Emsley L, Herrmann T, and Pintacuda G
- Subjects
- Protons, Nuclear Magnetic Resonance, Biomolecular methods, Proteins chemistry
- Abstract
Here we introduce a new pulse sequence for resonance assignment that halves the number of data sets required for sequential linking by directly correlating sequential amide resonances in a single diagonal-free spectrum. The method is demonstrated with both microcrystalline and sedimented deuterated proteins spinning at 60 and 111 kHz, and a fully protonated microcrystalline protein spinning at 111 kHz, with as little as 0.5 mg protein sample. We find that amide signals have a low chance of ambiguous linkage, which is further improved by linking in both forward and backward directions. The spectra obtained are amenable to automated resonance assignment using general-purpose software such as UNIO-MATCH.
- Published
- 2015
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