Rapid estimation of relative amide proton exchange rates of 15N‐labelled proteins by a straightforward water selective NOESY‐HSQC experiment

[摘要]

A straightforward heteronuclear pseudo-3D NOESY-HSQC pulse sequence using radiation damping to selectively invert magnetization at the water frequency was developed to estimate the amide proton exchange rates in ¹⁵N-labelled proteins. The peak intensities in the resultant 2D spectrum allow a direct classification of amide proton exchange rates according to short (ms), intermediate (ms to s) or long (≥ s) residence times. This method was successfully used for the analysis of amide proton exchange rates in the ¹⁵N-labelled FruR DNA-binding domain and pertinent information about its dynamics was obtained.

[学科分类] 生物化学/生物物理

[关键词] Hydrogen exchange rate;FruR;NMR;Protein dynamic;Radiation damping;Water-protein interaction;FruR;bacterial fructose repressor;FruR(1–57)∗;DNA-binding domain of FruR encompassing residues 1 to 57 and a LQHHHHHH C-terminal extension;NMR;nuclear magnetic resonance;NOE;nuclear Overhauser effect;NOESY;nuclear Overhauser enhancement spectroscopy;HSQC;1H-detected heteronuclear single quantum coherence spectroscopy;RD;radiation damping