Quantitative footprinting analysis. Binding to a single site.
The theory for measuring ligand binding constants from footprinting autoradiographic data associated with a single binding site is derived. If the ligand and DNA cleavage agent compete for a common site, the spot intensities are not proportional to the amount of DNA not blocked by ligand. The analysis of a single site is experimentally illustrated by using results for the anticancer drug actinomycin D interacting with the duplex d(TAGCGCTA)2 as probed with the hydrolytic enzyme DNase I.
Rehfuss
R
R
Department of Chemistry, Syracuse University, New York 13244-1200.
Goodisman
J
J
Dabrowiak
J C
JC
D001345:Autoradiography / Q000379:methods*; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism; D003609:Dactinomycin / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes / Q000378:metabolism
D008024:Ligands; D015345:Oligonucleotide Probes; D003609:Dactinomycin; D004247:DNA; D003850:Deoxyribonuclease I