Electrophoresis mobility shift assay and biosensor used in studying the interaction between bleomycin A5 and DNA.
DNA is the potential target of numerous drugs utilized widely in clinical cancer therapy. Here we employed bleomycin A5, with its deactivated form as contrast, to investigate the interaction between small pharmaceutical and DNA. Electrophoresis mobility shift assay (EMSA) which is a common technique used in studying specific interactions between DNA and proteins is applied in visualizing the binding of bleomycin A5 with DNA intuitively. More accurate association equilibrium constant for native and deactivated bleomycin A5 to DNA achieved on biosensor IAsys AUTO+ is 1.25x10(4) and 1.3x10(3) M(-1), respectively. With combination of EMSA and biosensor, a qualitative and quantitative method is described, which can be extended to studying the binding of most small molecules with targeting DNA and serves as a powerful tool in designing and screening for novel drugs.
Liu
Yanbin
Y
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
Wu
Fan
F
Zou
Guolin
G
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D001761:Bleomycin / Q000031:analogs & derivatives* / Q000737:chemistry; D002417:Cattle; D004247:DNA / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods*
D001761:Bleomycin; C025703:bleomycetin; D004247:DNA; C427203:calf thymus DNA