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

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 in:
Journal Of Biomolecular Nmr, 62, 3, 253-261
Year:
2015
Publisher:
Dordrecht, Springer Verlag
ISSN:
0925-2738
Keywords:
Laboratories:




 Record created 2015-09-28, last modified 2018-03-17


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