Investigation of the complexes of EcoRI endonuclease with decanucleotides containing canonical and modified recognition sequences using fluorescence and optical detection of magnetic resonance spectroscopy.
The binding of EcoRI endonuclease to the oligonucleotides d(GCGAATTCGC) and d(GCGAA) (5BrdU) (5BrdU) d(CGC) has been investigated to determine whether stacking interactions occur between tryptophan residues and the DNA bases. Fluorescence binding isotherms show that the decamer containing the canonical and that containing the modified recognition sequence bind with comparable affinity. Optically detected magnetic resonance spectra show limited perturbations of the Trp zero-field splitting parameters, which are assigned to electrical field effects. No evidence for Trp stacking interactions has been found.
Jhon
N I
NI
Department of Chemistry, University of California, Davis 95717.
Casas-Finet
J R
JR
Maki
A H
AH
Modrich
P
P
D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015246:Deoxyribonuclease EcoRI; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D014364:Tryptophan
D004268:DNA-Binding Proteins; D014364:Tryptophan; D004247:DNA; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI