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Old 06-17-2015, 09:27 PM
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Default Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.

Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.

Protein residue linking in a single spectrum for magic-angle spinning NMR assignment.

J Biomol NMR. 2015 Jun 16;

Authors: Andreas LB, Stanek J, Le Marchand T, Bertarello A, Paepe DC, Lalli D, Krej?íková M, Doyen C, Öster C, Knott B, Wegner S, Engelke F, Felli IC, Pierattelli R, Dixon NE, Emsley L, Herrmann T, Pintacuda G

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.


PMID: 26078089 [PubMed - as supplied by publisher]



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