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.
Macgregor
R B
RB
Department of Medicinal Chemistry, University of Illinois, Chicago 60612.
D001345:Autoradiography; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D016172:DNA Fingerprinting; D015246:Deoxyribonuclease EcoRI / Q000235:genetics* / Q000528:radiation effects; D006878:Hydroxides; D017665:Hydroxyl Radical; D008969:Molecular Sequence Data; D010957:Plasmids; D011487:Protein Conformation
D006878:Hydroxides; D017665:Hydroxyl Radical; D004247:DNA; D015246:Deoxyribonuclease EcoRI