A study of protein-water exchange through the off-resonance ROESY experiment: application to the DNA-binding domain of AlcR.
In this communication a new NMR experiment for the safe observation and quantification of water-protein exchange phenomena is presented. It combines a water-selective pulse, offering chemical shift-based separation, and the off-resonance ROESY dynamic filter, which permits the elimination of the unwanted intramolecular dipolar cross relaxation of protein protons. Moreover, pulsed field gradients are used for the suppression of radiation damping and the solvent signal. The straightforward incorporation of this sequence in heteronuclear experiments is demonstrated for the case of the DNA-binding domain of the alcohol regulator protein.
Birlirakis
N
N
Laboratoire de RMN, ICSN-CNRS, Gif-sur-Yvette, France.
Cerdan
R
R
Guittet
E
E
D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000737:chemistry; D001233:Aspergillus nidulans / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D002417:Cattle; D004271:DNA, Fungal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005069:Evaluation Studies as Topic; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D015394:Molecular Structure; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D014867:Water
C074202:ALCR protein, Aspergillus nidulans; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D014157:Transcription Factors; D014867:Water; D007611:Aprotinin