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The interaction of 7-aminoactinomycin (7AAMD) with selected DNA sequences (TAGTTA, R5, HP5, and HP1) of different lengths and secondary structures, all containing a 5'-TAGT-3' block, was studied at an ionic strength of 0.02 M and pH 7.7 by means of fluorescence equilibrium and kinetic (stopped-flow) measurements. Both approaches indicated that the antibiotic binds strongly to both the single-stranded and hairpin (HP1) structures, although the sequences lacked the canonical GpC sites favored by actinomycin. Binding isotherms and initial rate analyses revealed that the binding stoichiometry was 1:1 in all cases. While the single-stranded sequences displayed a simple monoexponential kinetic behavior, the binding of 7AAMD to HP1 at <30°C was biphasic and could be rationalized in terms of a sequential formation of two isomeric bound forms or alternatively in terms of an ssHP1-hpHP1 equilibrium, with both HP1 forms reacting with 7AAMD. The rates of complex dissociation induced by the detergent SDS were also measured. After correction of the kinetic traces for spurious effects that can be attributed to the SDS, monoexponential traces were obtained, with relaxation times in agreement with the kinetics of complex formation. © 2009 American Chemical Society.


Documento: Artículo
Título:7-aminoactinomycin binding to DNA sequences lacking GpC sites: A thermodynamic and kinetic study
Autor:Biver, T.; Venturini, M.; Jares-Erijman, E.A.; Jovin, T.M.; Secco, F.
Filiación:Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Risorgimento 35, 56126 Pisa, Italy
Departamento de Qúimica Orgánica, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria Pabellón II/3, 1428 Buenos Aires, Argentina
Laboratory of Cellular Dynamics, Max Planck Institute for Biophysical Chemistry, am Fassberg 11, 37077 Göttingen, Germany
Palabras clave:Binding isotherms; Complex dissociation; Complex formations; Initial rate; Secondary structures; Atmospheric temperature; Diffusers (optical); DNA; DNA sequences; Genes; Ionic strength; Nucleic acids; Organic acids; Stoichiometry; Binding sites; 7 aminodactinomycin; hairpin DNA; single stranded DNA; unclassified drug; article; binding affinity; binding assay; binding kinetics; complex formation; dissociation; DNA sequence; DNA structure; drug DNA binding; drug DNA interaction; equilibrium constant; fluorescence spectroscopy; molecular stability; priority journal; stoichiometry; thermodynamics; Base Sequence; Dactinomycin; DNA; Fluorescent Dyes; Kinetics; Nucleic Acid Conformation; Thermodynamics
Página de inicio:173
Página de fin:179
Título revista:Biochemistry
Título revista abreviado:Biochemistry
CAS:7 aminodactinomycin, 7240-37-1; 7-aminoactinomycin D, 7240-37-1; DNA, 9007-49-2; Dactinomycin, 50-76-0; Fluorescent Dyes


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---------- APA ----------
Biver, T., Venturini, M., Jares-Erijman, E.A., Jovin, T.M. & Secco, F. (2009) . 7-aminoactinomycin binding to DNA sequences lacking GpC sites: A thermodynamic and kinetic study. Biochemistry, 48(1), 173-179.
---------- CHICAGO ----------
Biver, T., Venturini, M., Jares-Erijman, E.A., Jovin, T.M., Secco, F. "7-aminoactinomycin binding to DNA sequences lacking GpC sites: A thermodynamic and kinetic study" . Biochemistry 48, no. 1 (2009) : 173-179.
---------- MLA ----------
Biver, T., Venturini, M., Jares-Erijman, E.A., Jovin, T.M., Secco, F. "7-aminoactinomycin binding to DNA sequences lacking GpC sites: A thermodynamic and kinetic study" . Biochemistry, vol. 48, no. 1, 2009, pp. 173-179.
---------- VANCOUVER ----------
Biver, T., Venturini, M., Jares-Erijman, E.A., Jovin, T.M., Secco, F. 7-aminoactinomycin binding to DNA sequences lacking GpC sites: A thermodynamic and kinetic study. Biochemistry. 2009;48(1):173-179.