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A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins
Author(s) -
Wiedemann Christoph,
Bellstedt Peter,
Häfner Sabine,
Herbst Christian,
Bordusa Frank,
Görlach Matthias,
Ohlenschläger Oliver,
Ramachandran Ramadurai
Publication year - 2016
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.201600155
Subject(s) - heteronuclear molecule , chemistry , magnetization transfer , amide , magnetization , chemical shift , pulse sequence , two dimensional nuclear magnetic resonance spectroscopy , spectral line , nuclear magnetic resonance , heteronuclear single quantum coherence spectroscopy , resonance (particle physics) , nuclear magnetic resonance spectroscopy , analytical chemistry (journal) , stereochemistry , physics , magnetic field , magnetic resonance imaging , atomic physics , organic chemistry , medicine , quantum mechanics , astronomy , radiology
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 α 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 J CαN and 2 J CαN 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 α‐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.