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.

Protein residue linking in a single spectrum for magic-angle spinning NMR assignment / Andreas, Loren B.; Stanek, Jan; Le Marchand, Tanguy; Bertarello, Andrea; Cala-De Paepe, Diane; Lalli, Daniela; Krejčíková, Magdaléna; Doyen, Camille; Öster, Carl; Knott, Benno; Wegner, Sebastian; Engelke, Frank; Felli, Isabella C.; Pierattelli, Roberta; Dixon, Nicholas E.; Emsley, Lyndon; Herrmann, Torsten; Pintacuda, Guido. - In: JOURNAL OF BIOMOLECULAR NMR. - ISSN 0925-2738. - STAMPA. - 62:(2015), pp. 253-261. [10.1007/s10858-015-9956-1]

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

LALLI, DANIELA;FELLI, ISABELLA CATERINA;PIERATTELLI, ROBERTA;
2015

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.
2015
62
253
261
Andreas, Loren B.; Stanek, Jan; Le Marchand, Tanguy; Bertarello, Andrea; Cala-De Paepe, Diane; Lalli, Daniela; Krejčíková, Magdaléna; Doyen, Camille; ...espandi
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1041723
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