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., Goodisman, J., & Dabrowiak, J. C. (1990). Quantitative footprinting analysis. binding to a single site. Biochemistry, 29(3), 777-781.
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