Thermodynamics and kinetics of co-operative protein-nucleic acid binding: II. Studies on the binding between protamine and calf thymus DNA |
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Authors: | Fumiyuki Watanabe Gerhard Schwarz |
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Institution: | Department of Biophysical Chemistry Biocenter, University of Basel CH-4056 Basel, Switzerland |
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Abstract: | Unspecific binding of a protamine, namely fluorescein-labelled clupeine Z, to double-stranded calf thymus DNA was studied using fluorescence titration methods and chemical relaxation techniques. Both equilibrium and kinetic data have been analysed using general theoretical approaches discussed in the accompanying paper. The results agree well with the predictions made on the basis of a standard co-operative binding model.Basic parameters evaluated are the co-operative binding constant (K), the coefficient measuring co-operative interaction between nearest neighbours (q), the number of nucleotides occupied by one protamine molecule (n) and the rate constant of dissociation at the ends of bound ligand sequences (KD). Values obtained at 20 °C, pH 7.5 and 0.4 m-NaCl were K = 5.8 × 107m?1, q = 1700, n = 20 and KD = 0.29 s?1. They have been found to be sensitive to the concentration of added salt (NaCl). This effect apparently reflects the essentially electrostatic nature of the binding process. The results can be satisfactorily described in terms of competitive binding of sodium ions. |
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Keywords: | Author to whom all correspondence should be addressed |
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