G. Bischoff et al., DNA-DRUG INTERACTION MEASUREMENTS USING SURFACE-PLASMON RESONANCE, Journal of biomolecular structure & dynamics, 16(2), 1998, pp. 187-203
The interactions of the drugs 2,7-bis[(diethylamino)-ethoxy]-fluoren-9
-one dihydrochloride (<(Tilorone)under bar>), 2,7-bis[(dipropylamino)-
acetamido]-fluoren-9-one dihydrochloride (<(FA-2)under bar>), -(4-meth
yl-1-piperazinyl)-2,5'-bi-1H-benzimidazole trihydrochloride (<(Hoechst
33258)under bar>), and hematoporphyrin IX derivative (<(HPD)under bar
>) with synthetic self-complementary DNA (36-b.p.; 5'-biotin-spacer-[d
(CGCTATATAGCG)](3)-3') were studied by SPR (Surface Plasmon Resonance)
. Monolayers of biotinylated DNA were immobilized on a streptavidin-de
xtran-gold triple-layer. Small portions of the drugs (approximately 30
pmol/ml) were injected in continuous flow. The mass corresponded to t
he amount of the bound molecules. Injections of 50 mM sodium hydroxide
pulses separated the DNA double strands, releasing the effector molec
ules. Subsequent treatments with the effecters gave reproducible resul
ts. The maximum interaction between drug and DNA was observed in the c
ase of Tilorone. 41 molecules could bind to the 36-b.p. DNA duplex. To
investigate the microscopic behavior in condensed nucleic acid phases
, SFM (Scanning Force Microscopy)-imaging and polarizing microscopic o
bservations of DNA-effector complexes were carried out. Supplementary
UV-absorption thermal denaturation curves of DNA with the above-mentio
ned effecters in dilute solutions were measured. As an additional aid
to understand the geometries of DNA-drug interactions, computer simula
tions were performed and compared with the experimental data.