Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides

C Behrens; N Harrit; P E Nielsen

Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides

C Behrens, N Harrit, P E Nielsen

37 Citations (Scopus)

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.

Original language	English
Journal	Bioconjugate Chemistry
Volume	12
Issue number	6
Pages (from-to)	1021-7
Number of pages	7
ISSN	1043-1802
Publication status	Published - 22 Nov 2001

Keywords

Affinity Labels
Amino Acid Motifs
Amino Acid Sequence
Amino Acids
Animals
Benzimidazoles
DNA
DNA Footprinting
DNA-Binding Proteins
Esters
Fluorescent Dyes
Heterocyclic Compounds, 1-Ring
Histones
Humans
Intercalating Agents
Peptide Fragments
Spectrum Analysis

Cite this

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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.",

keywords = "Affinity Labels, Amino Acid Motifs, Amino Acid Sequence, Amino Acids, Animals, Benzimidazoles, DNA, DNA Footprinting, DNA-Binding Proteins, Esters, Fluorescent Dyes, Heterocyclic Compounds, 1-Ring, Histones, Humans, Intercalating Agents, Peptide Fragments, Spectrum Analysis",

author = "C Behrens and N Harrit and Nielsen, {P E}",

year = "2001",

month = nov,

day = "22",

language = "English",

volume = "12",

pages = "1021--7",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

number = "6",

}

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

SN - 1043-1802

VL - 12

SP - 1021

EP - 1027

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 6

ER -

Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides

Abstract

Keywords

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