Footprinting of EcoRI endonuclease at high pressure
Copyright policy )Footprinting of EcoRI endonuclease at high pressure
Hydroxyl radicals generated by irradiation with gamma rays have been used to footprint EcoRI endonuclease with single base pair resolution at pressures up to 144 MPa. At atmospheric pressure (0.1 MPa) a 10 base pair footprint was found. With increasing pressure three types of responses were observed: (1) bases distant from the recognition sequence showed a moderate increase in solvent exposure; (2) the bases at the point of enzymatic activity showed a large increase in cleavage by the hydroxyl radicals; and (3) the two center-most bases exhibited no pressure-induced change in solvent accessibility. The results are interpreted in terms of localized conformational changes of EcoRI.
- University of Illinois at Chicago United States
Base Sequence, Hydroxyl Radical, Protein Conformation, Molecular Sequence Data, Hydroxides, Autoradiography, DNA, DNA Fingerprinting, Deoxyribonuclease EcoRI, Plasmids
Base Sequence, Hydroxyl Radical, Protein Conformation, Molecular Sequence Data, Hydroxides, Autoradiography, DNA, DNA Fingerprinting, Deoxyribonuclease EcoRI, Plasmids
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