Use of capillary electrophoresis in the study of ligand-DNA interactions | Zendy

Imad I. Hamdan | Zendy; G.G. Skellern | Zendy; R. D. WAIGH | Zendy

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Use of capillary electrophoresis in the study of ligand-DNA interactions

Author(s) -

Imad I. Hamdan,

G.G. Skellern,

R. D. WAIGH

Publication year - 1998

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/26.12.3053

Subject(s) - netropsin , ethidium bromide , cooperativity , capillary electrophoresis , oligonucleotide , biology , dna , cooperative binding , ligand (biochemistry) , intercalation (chemistry) , binding site , binding selectivity , biophysics , microbiology and biotechnology , biochemistry , minor groove , chemistry , receptor , inorganic chemistry

Free solution capillary electrophoresis (FSCE) has been used to separate two non-self-complementary 12mer oligonucleotide duplexes: d(AAATTATATTAT).d(ATAA-TATAATTT) and d(GGGCCGCGCCGC).d(GCGGCGCGGCCC). Titration of mixtures of the two oligonucleotides with model intercalators (ethidium bromide andactinomycin D) and minor groove binders (netropsin, Hoechst 33258 and distamycin) has shown the suitability of FSCE as a method to study the sequence selectivity of DNA binding agents. Binding data have shown cooperativity of binding for netropsin and Hoechst 33258 and have provided ligand:DNA binding ratios for all five compounds. Cooperativity of netropsin binding to a 12mer with two potential sites has been demonstrated for the first time. Ligands binding in the minor groove caused changes in migration time and peak shape which were significantly different from those caused by intercalators.

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