1. Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.
- Author
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Andreas, Loren, Stanek, Jan, Marchand, Tanguy, Bertarello, Andrea, Paepe, Diane, Lalli, Daniela, Krejčíková, Magdaléna, Doyen, Camille, Öster, Carl, Knott, Benno, Wegner, Sebastian, Engelke, Frank, Felli, Isabella, Pierattelli, Roberta, Dixon, Nicholas, Emsley, Lyndon, Herrmann, Torsten, and Pintacuda, Guido
- 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. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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