Optimization of cross-linked lexitropsins.
In attempts to optimize the cross-linked lexitropsin design, a number of cross-linked dimers composed of two tris(N-methylpyrrolecarboxamide) strands were synthesized and their binding interactions with poly d(A).poly d(T) and poly d(A-T).poly d(A-T) were characterized by circular dichroism and ethidium fluorometry. While all alkanediyl-linked dimers showed a similar binding behavior to the homo AT polymer, particularly at low ligand concentrations, the decanediyl linker was found to be the optimal linker permitting the bidentate antiparallel side-by-side binding of the corresponding dimer to the alternating AT polymer. Thus, in comparison with the monomer, the decanediyl-linked dimer has a binding strength enhancement of about 1400 times in the 1:1 binding mode. Moreover, the hydrophilicity of the linker dimer has a significant effect on the bidentate binding strength. The (3,6)-dioxaoctanediyl-linked dimer has a further binding strength enhancement of 10 times over the decanediyl-linked dimer. Overall, the best optimized dimer has a binding strength enhancement of over 14,000 times in comparison with the monomer in the 1:1 binding mode. This binding enhancement parallels that observed in the best optimized bisintercalators. Distance-restrained molecular modeling provides support for the experimental results. Dimers of longer linkers can readily accommodate a bidentate antiparallel side-by-side binding mode but those of shorter linkers necessitate marked structural distortions in the bound ligand molecules. It is further observed that the binding strength enhancement to the alternating AT polymer is not always accompanied by the binding specificity improvement. Our analysis suggests that the non-specific appendage-DNA backbone interaction is a key factor that controls the specificity improvement.
Chen
Y H
YH
Department of Chemistry, University of Alberta, Edmonton, Canada.
Yang
Y
Y
Lown
J W
JW
D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000378:metabolism; D004996:Ethidium / Q000737:chemistry; D005470:Fluorometry / Q000379:methods; D008956:Models, Chemical; D008958:Models, Molecular; D009429:Netropsin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D017186:Titrimetry
D003432:Cross-Linking Reagents; D009841:Oligonucleotides; C053654:lexitropsin; D009429:Netropsin; D004996:Ethidium