DNA-DRUG INTERACTION MEASUREMENTS USING SURFACE-PLASMON RESONANCE

Citation
G. Bischoff et al., DNA-DRUG INTERACTION MEASUREMENTS USING SURFACE-PLASMON RESONANCE, Journal of biomolecular structure & dynamics, 16(2), 1998, pp. 187-203
Citations number
50
Categorie Soggetti
Biophysics,Biology
ISSN journal
07391102
Volume
16
Issue
2
Year of publication
1998
Pages
187 - 203
Database
ISI
SICI code
0739-1102(1998)16:2<187:DIMUSR>2.0.ZU;2-N
Abstract
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.