A quantitative assay to measure the relative DNA-binding affinity of pyrrolo[2,1-c] [1,4]benzodiazepine (PBD) antitumour antibiotics based on the inhibition of restriction endonuclease BamHI.
An assay has been developed (restriction endonuclease digestion assay--RED100) based on inhibition of the restriction endonuclease BamHI that is capable of quantitative evaluation of the relative DNA-binding affinity of pyrrolo[2,1-c] [1,4]benzodiazepine (PBD) antitumour antibiotics. This method provides comparable results to those obtained from thermal denaturation and ethidium bromide displacement assays but is much more sensitive, discriminating between molecules of similar structure such as DC-81, iso-DC-81 and neothramycin. The results reveal a trend between relative DNA-binding affinity and in vitro cytotoxicity for the PBDs in two tumour cell lines studied.
Puvvada
M S
MS
School of Pharmacy and Biomedical Sciences, University of Portsmouth, London, UK.
Hartley
J A
JA
Jenkins
T C
TC
Thurston
D E
DE
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D001483:Base Sequence; D001570:Benzodiazepinones / Q000378:metabolism*; D002417:Cattle; D004247:DNA / Q000378:metabolism*; D015245:Deoxyribonuclease BamHI / Q000037:antagonists & inhibitors*; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011485:Protein Binding
D000903:Antibiotics, Antineoplastic; D001570:Benzodiazepinones; D004247:DNA; D015245:Deoxyribonuclease BamHI