Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides
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Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides. / Behrens, C; Harrit, N; Nielsen, P E.
In: Bioconjugate Chemistry, Vol. 12, No. 6, 22.11.2001, p. 1021-7.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides
AU - Behrens, C
AU - Harrit, N
AU - Nielsen, P E
PY - 2001/11/22
Y1 - 2001/11/22
N2 - 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.
AB - 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.
KW - Affinity Labels
KW - Amino Acid Motifs
KW - Amino Acid Sequence
KW - Amino Acids
KW - Animals
KW - Benzimidazoles
KW - DNA
KW - DNA Footprinting
KW - DNA-Binding Proteins
KW - Esters
KW - Fluorescent Dyes
KW - Heterocyclic Compounds, 1-Ring
KW - Histones
KW - Humans
KW - Intercalating Agents
KW - Peptide Fragments
KW - Spectrum Analysis
M3 - Journal article
C2 - 11716695
VL - 12
SP - 1021
EP - 1027
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
SN - 1043-1802
IS - 6
ER -
ID: 154517776