ausblenden:
Schlagwörter:
NMR; Intrinsically disordered protein; Assignment;Alpha-synuclein; APSY; Solvent exchange
Zusammenfassung:
Sequence specific resonance assignment is the prerequisite for the NMR-based analysis of the conformational ensembles and their underlying dynamics of intrinsically disordered proteins. However, rapid solvent exchange in intrinsically disordered proteins often complicates assignment strategies based on HN-detection. Here we present a six-dimensional alpha proton detection-based automated projection spectroscopy (APSY) experiment for backbone assignment of intrinsically disordered proteins. The 6D HCACONCAH APSY correlates the six different chemical shifts, Halpha(i-1), Calpha(i-1), C'(i-1), N(i), Calpha(i) and Halpha(i). Application to two intrinsically disordered proteins, 140-residue alpha-synuclein and a 352-residue isoform of Tau, demonstrates that the chemical shift information provided by the 6D HCACONCAH APSY allows efficient backbone resonance assignment of intrinsically disordered proteins.
