A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins.
 

A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins.

A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins.

Chemphyschem. 2016 Apr 9;

Authors: Wiedemann C, Bellstedt P, Häfner S, Herbst C, Bordusa F, Görlach M, Ohlenschläger O, Ramachandran R

Abstract
The RF pulse scheme RN[N-CA HEHAHA]NH, which provides a convenient approach to the acquisition of different multidimensional chemical shift correlation NMR spectra leading to backbone resonance assignments, including those of the proline residues of intrinsically disordered proteins (IDPs), is experimentally demonstrated. Depending on the type of correlation data required, the method involves the generation of in-phase ((15) N)(x) magnetisation via different magnetisation transfer pathways such as H->N->CO->N, HA->CA->CO->N, H->N->CA->N and H->CA->N, the subsequent application of (15) N-(13) C(?) heteronuclear Hartmann-Hahn mixing over a period of ?100 ms, chemical-shift labelling of relevant nuclei before and after the heteronuclear mixing step and amide proton detection in the acquisition dimension. It makes use of the favourable relaxation properties of IDPs and the presence of (1) JC?N and (2) JC?N couplings to achieve efficient correlation of the backbone resonances of each amino acid residue "i" with the backbone amide resonances of residues "i-1" and "i+1". It can be implemented in a straightforward way through simple modifications of the RF pulse schemes commonly employed in protein NMR studies. The efficacy of the approach is demonstrated using a uniformly ((15) N,(13) C) labelled sample of ?-synuclein. The different possibilities for obtaining the amino-acid-type information, simultaneously with the connectivity data between the backbone resonances of sequentially neighbouring residues, have also been outlined.


PMID: 27061973 [PubMed - as supplied by publisher]



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