A six-dimensional alpha proton detection-based APSY experiment for backbone assignment of intrinsically disordered proteins
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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, Hα(i − 1), Cα(i − 1), C′(i − 1), N(i), Cα(i) and Hα(i). Application to two intrinsically disordered proteins, 140-residue α-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.
KeywordsNMR Intrinsically disordered protein Assignment α-Synuclein APSY Solvent exchange
We thank Eckhard Mandelkow and Jacek Biernat for the htau23 sample. This work was in part supported by the DFG through ZW71/3-2 and ZW71/7-1.
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