NMR characterization of the DNA binding properties of a novel Hoechst 33258 analogue peptide building block.
A novel aryl-bis-benzimidazole amino acid analogue of the DNA-binding compound Hoechst 33258 has recently been designed for incorporation in peptide combinatorial libraries by replacing the N-methylpiperazine group with a carboxyl group and the hydroxy group with an amino-methyl group. The DNA-binding properties of the aryl-bis-benzimidazole monomer with the C-terminus derivatized with 3-(dimethylamino)-propylamine has been investigated in this paper by (1)H NMR studies of two different complexes with two different DNA sequences: A(5) d(5'-GCCA(5)CG-3'):d(5'-CGT(5)GGC-3') and A(3)T(3) d(5'-CGA(3)T(3)CG-3')(2). Chemical shift footprinting shows that the ligand binds at the center of the A(3)T(3) sequence but at the 3'-end of A(5). A large number of NOEs show a well-defined complex with the ligand situated at the center of the palindromic A(3)T(3) but with the asymmetric A(5) the ligand binds with an orientational preference with the bis-benzimidazole moiety displaced toward the 3'-end from the center of the duplex. Two families of models of the complexes with A(5) and A(3)T(3) were derived with restrained molecular dynamics based on a large set of 70 and 61, respectively, intermolecular ligand NOEs. Both models give a picture of a tightly fitting ligand with close van der Waals contacts with the walls of the minor groove and with the two benzimidazole and the amide hydrogens involved in bifurcated cross-strand hydrogen bonds to adenine N3 and thymine O2. The minor groove width of the models correlate well with the binding site of the ligand, and the orientational preference is argued to be a consequence of the minor groove width and hydrogen bonding.
Bunkenborg
Jakob
J
Department of Chemistry, University of Southern Denmark, Odense University, Odense M, DK-5230 Denmark.
Behrens
Carsten
C
Jacobsen
Jens Peter
JP
D001665:Binding Sites; D006690:Bisbenzimidazole / Q000031:analogs & derivatives*; D004247:DNA / Q000378:metabolism*; D006860:Hydrogen Bonding; D008024:Ligands; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009841:Oligonucleotides / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D013329:Structure-Activity Relationship
D008024:Ligands; D009841:Oligonucleotides; D009842:Oligopeptides; D004247:DNA; D006690:Bisbenzimidazole