title: Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides.
abstract: The synthesis of a new versatile "Hoechst 33258-like" Boc-protected amino acid building block for peptide synthesis is described. It is demonstrated that this new ligand is an effective mimic of Hoechst 33258 in terms of DNA affinity and sequence specificity. Furthermore, this minor groove binder was conjugated to a DNA condensing peptide (KSPKKAKK) by continuous solid-phase peptide synthesis, and the conjugate exhibited increased DNA affinity (ca. 10-fold), but similar sequence preference compared to Hoechst 33258 as analyzed by DNaseI footprinting. Finally, the fluorescence quantum yield of the new chromophore is found to increase 30% upon binding to double stranded DNA.
authors.affiliation: Department of Chemistry, University of Copenhagen, Universitetsparken 5, DK-2100, Copenhagen, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Behrens
authors.forename: N
authors.initials: N
authors.lastname: Harrit
authors.forename: P E
authors.initials: PE
authors.lastname: Nielsen
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000737:chemistry; D004952:Esters / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D006573:Heterocyclic Compounds, 1-Ring / Q000737:chemistry*; D006657:Histones; D006801:Humans; D007364:Intercalating Agents / Q000138:chemical synthesis* / Q000378:metabolism; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000378:metabolism; D013057:Spectrum Analysis
chemical_list: D000345:Affinity Labels; D000596:Amino Acids; D001562:Benzimidazoles; D004268:DNA-Binding Proteins; C447681:DOTA tris(phenylmethyl)ester; D004952:Esters; D005456:Fluorescent Dyes; D006573:Heterocyclic Compounds, 1-Ring; D006657:Histones; D007364:Intercalating Agents; D010446:Peptide Fragments; D004247:DNA