Thermal denaturation profiles of deoxypolynucleotide-ligand complexes: semiempirical studies.
We have semiempirically studied the thermal denaturation profiles of complexes formed between double strand polynucleotides and pure stabilizer nonspecific binding ligands. By using the McGhee model (J. D. McGhee, (1976) Biopolymers 15, 1345-1375) we have found a simple, analytical relationship between the melting temperature (Tm) and the Kh (intrinsic association constant), nh (apparent site size), and wh (cooperativity constant) values of the interaction. The validity of this approach strongly depends on the sigma value (sigma being the nucleation parameter of the DNA). Through the equation so obtained it is possible to calculate the Kh, nh, and wh values from the melting temperature of three experimental thermal denaturation profiles at different r (ligand/polynucleotide ratio) values. The method has been checked by studying the thermal denaturation profiles of daunomycin-poly(d(A-T)).poly(d(A-T)) complexes in two different salt concentrations. The results so obtained are compared with those previously described using other techniques. The applicability of the method here developed is discussed in relation with both the nature of the ligands and the value of the nucleation parameter (sigma).
Rodriguez
A T
AT
Departamento de Bioquimica y Biologia Molecular I, Facultad de Ciencias Quimicas, Universidad Complutense, Madrid, Spain.
Fernandez
B A
BA
Montero
F
F
D004247:DNA / Q000648:ultrastructure; D003630:Daunorubicin; D006358:Hot Temperature; D008024:Ligands; D009691:Nucleic Acid Denaturation
D008024:Ligands; D004247:DNA; D003630:Daunorubicin