Rapid estimation of relative amide proton exchange rates of 15 N-labelled proteins by a straightforward water selective NOESY-HSQC experiment.
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 (> or = 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.
Böckmann
A
A
Laboratoire de RMN, ISCN-CNRS, Gif sur Yvette, France.
Penin
F
F
Guittet
E
E
D000577:Amides; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D009690:Nucleic Acid Conformation; D011336:Probability; D011522:Protons; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism; D056968:Lac Repressors
D000577:Amides; D001426:Bacterial Proteins; D004247:DNA; D004268:DNA-Binding Proteins; D009587:Nitrogen Isotopes; D011522:Protons; D012097:Repressor Proteins; C068837:FruR protein, Bacteria; D056968:Lac Repressors