ABSTRACT
Thermodynamic parameters of melting process (DeltaHm, Tm, DeltaTm) of calf thymus DNA, poly(dA)poly(dT) and poly(d(A-C)).poly(d(G-T)) were determined in the presence of various concentrations of TOEPyP(4) and its Zn complex. The investigated porphyrins caused serious stabilization of calf thymus DNA and poly poly(dA)poly(dT), but not poly(d(A-C))poly(d(G-T)). It was shown that TOEpyp(4) revealed GC specificity, it increased Tm of satellite fraction by 24 degrees C, but ZnTOEpyp(4), on the contrary, predominantly bound with AT-rich sites and increased DNA main stage Tm by 18 degrees C, and Tm of poly(dA)poly(dT) increased by 40 degrees C, in comparison with the same polymers without porphyrin. ZnTOEpyp(4) binds with DNA and poly(dA)poly(dT) in two modes--strong and weak ones. In the range of r from 0.005 to 0.08 both modes were fulfilled, and in the range of r from 0.165 to 0.25 only one mode--strong binding--took place. The weak binding is characterized with shifting of Tm by some grades, and for the strong binding Tm shifts by approximately 30-40 degrees C. Invariability of DeltaHm of DNA and poly(dA)poly(dT), and sharp increase of Tm in the range of r from 0.08 to 0.25 for thymus DNA and 0.01-0.2 for poly(dA)poly(dT) we interpret as entropic character of these complexes melting. It was suggested that this entropic character of melting is connected with forcing out of H2O molecules from AT sites by ZnTOEpyp(4) and with formation of outside stacking at the sites of binding. Four-fold decrease of calf thymus DNA melting range width DeltaTm caused by increase of added ZnTOEpyp(4) concentration is explained by rapprochement of AT and GC pairs thermal stability, and it is in agreement with a well-known dependence, according to which DeltaT approximately TGC-TAT for DNA obtained from higher organisms (L. V. Berestetskaya, M. D. Frank-Kamenetskii, and Yu. S. Lazurkin. Biopolymers 13, 193-205 (1974)). Poly (d(A-C))poly(d(G-T)) in the presence of ZnTOEpyp(4) gives only one mode of weak binding. The conclusion is that binding of ZnTOEpyp(4) with DNA depends on its nucleotide sequence.
Subject(s)
DNA/metabolism , Poly dA-dT/metabolism , Polydeoxyribonucleotides/metabolism , Porphyrins/metabolism , Pyridinium Compounds/metabolism , Zinc/metabolism , Animals , Calorimetry , Cattle , DNA/chemistry , Hot Temperature , Nucleic Acid Denaturation , Poly dA-dT/chemistry , Polydeoxyribonucleotides/chemistry , Porphyrins/chemistry , Pyridinium Compounds/chemistry , Zinc/chemistryABSTRACT
We employed UV light absorption and circular dichroism (CD) spectroscopic measurements to study the binding of novel water-soluble porphyrins meso-tetra-(4N-allylpyridyl)porphyrin [TAlPyP4], and its Ag containing derivative to the poly(rA)poly(rU) and poly(rI)poly(rC) RNA duplexes. Our results suggest that TAlPyP4 associate with the duplexes via intercalation, whereas the conservative CD spectra indicates that AgTAlPyP4 preferably binds via outside self-stacking mode. We used our determined binding isotherms for each ligand-RNA binding event to calculate the binding constant, Kb, and binding free energy, DeltaGb = -RTlnKb. By performing these experiments as a function of temperature, we evaluated the van't Hoff binding enthalpies, DeltaH. The binding entropies, DeltaSb, were calculated as DeltaSb = (DeltaHb - DeltaGb)/T. We interpret our data in terms of specific interactions that stabilize/destabilize each ligand-RNA complex studied in this work. Taken together, our data provide important new information about the thermodynamics of interactions of porphyrins with nucleic acids.
Subject(s)
Poly A-U/metabolism , Poly I-C/metabolism , Porphyrins/metabolism , Thermodynamics , Poly A-U/chemistry , Poly I-C/chemistry , Porphyrins/chemistryABSTRACT
The peculiarities of helix-coil transition of DNA at complex-formation with cis-diamminedichlorplatinum (II) (cis-DDP) have been investigated in acidic environment by UV absorbance technique. It is shown that observed features of behavior T (pH) and T(m) (pH) DNA at pH 2.8-3.0 are possible to explain by formation in DNA of pseudo-ring structures at covalent linking of cis-DDP with DNA.
Subject(s)
Cisplatin/chemistry , Cisplatin/metabolism , DNA/chemistry , DNA/metabolism , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/metabolism , Cattle , Hydrogen-Ion Concentration , Nucleic Acid Denaturation , Transition TemperatureABSTRACT
Interactions of meso-tetra-(4-N-oxyethylpyridyl) porphyrin (TOEPyP(4)), its 3-N analog (TOEPyP(3)) and their Co, Cu, Ni, Zn metallocomplexes with duplex DNA have been investigated by uv/visible absorbance and circular dichrosim spectroscopies. Results reveal the interactions of these complexes with duplex DNA are of two types. (1) External binding of duplex DNA by metalloporphyrins containing Zn and Co, and (2) Binding of duplex DNA both externally and internally (by intercalation) by porphyrins not containing metals, and metalloporphyrins containing Cu and Ni. Results indicate that (4N-oxyethylpyridyl) porphyrins intercalate more preferably in the structure of duplex DNA and have weaker external binding than 3N-porphyrins.
Subject(s)
DNA/chemistry , Porphyrins/chemistry , Circular Dichroism , Metals , Nucleic Acid Heteroduplexes , Spectrophotometry, Ultraviolet/methodsABSTRACT
The complex formation of porphyrins with DNA leads to changes of stability of DNA. In the present study we investigated binding properties and the thermodynamic parameters of a water-soluble, cationic planar Cu(II)-containing meso-tetrakis(4-N-butyl-pyridiniumyl)porphyrin [CuTButPyP4] and nonplanar Co(II)-containing meso-tetrakis(4-N-butyl-pyridiniumyl)porphyrin [CoButPyP4] with calf thymus DNA in the presence of divalent manganese ions. For displaying the changes of thermodynamic parameters (T(m) and ΔT) the melting curves of DNA-porphyrin complexes in the presence of Mn(2+) ions have been obtained. The enthalpy (ΔH) of helix-coil transition has been also evaluated. It was shown that the binding of ions to DNA proceeds in two stages depending on the manganese/DNA phosphates molar ratio [Mn]/[P]. At the first stage (0.001<[Mn]/[P]<1), the interaction of manganese ions with DNA phosphates occurs, causing an additional screening of their negative charge and the stabilization of the double helix. As a result, the best conditions for intercalation of CuTButPyP4 or of peripheral rings of CoButPyP4 occur. The significant increase of T(m), but less changes of ΔT were observed. At the second stage (1<[Mn]/[P]<4), the ions interact with both the phosphates and the nitrogen bases of DNA. At this stage, it is possible for the manganese ion to coordinate simultaneously to the oxygen atom of the phosphate and the neighboring base of DNA. At a higher [Mn]/[P] ratio, the destabilization of the double helix begins, and partial breakage of the hydrogen bonds between the nitrogen bases occurs. Respectively the destabilization of DNA in the presence of both porphyrins takes place.
Subject(s)
DNA/metabolism , Manganese/metabolism , Porphyrins/metabolism , Animals , Cattle , Circular Dichroism , DNA/chemistry , Ions/chemistry , Ions/metabolism , Manganese/chemistry , Models, Molecular , Nucleic Acid Denaturation , Porphyrins/chemistry , ThermodynamicsABSTRACT
The interaction of meso-tetra-(4N-oxyethylpyridyl)porphyrin (TOEPyP4) and its Zn(II)-, Cu(II)-, Mn(III)-derivatives with tRNA from E.Coli at low ionic strength (micro=0.02M) was studied using UV/Vis spectrophotometry and Circular Dichroism (CD) methods. An unusual Induced Circular Dichroism (ICD) spectra profile of the ZnTOEPyP4-tRNA complex is found. It is demonstrated that ZnTOEPyP4 is ordered in a stack, not only on helical sites, but also on loops of a hairpin form of tRNA. TOEPyP4 and CuTOEPyP4 are able to intercalate in the helical sites of this form of tRNA. MnTOEPyP4 interacts with tRNA via external non-ordered mechanism. It is established that all porphyrins are bound with tRNA more strongly than with DNA.