Title

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

RIS ID

101570

Publication Details

Andreas, L. B., Stanek, J., Le Marchand, T., Bertarello, A., Paepe, D. Cala-De., Lalli, D., Krejcikova, M., Doyen, C., Öster, C., Knott, B., Wegner, S., Engelke, F., Felli, I. C., Pierattelli, R., Dixon, N. E., Emsley, L., Herrmann, T. & Pintacuda, G. (2015). Protein residue linking in a single spectrum for magic-angle spinning NMR assignment. Journal of Biomolecular NMR, 62 (3), 253-261.

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.

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Link to publisher version (DOI)

http://dx.doi.org/10.1007/s10858-015-9956-1