Vladimir A. Valeev

Adsorption of Poly(rA) on the Carbon Nanotube Surface and its Hybridization with Poly(rU)

Adsorption of poly(rA) on a single-walled carbon nanotube surface in aqueous suspension and the subsequent hybridization of this polymer with free poly(rU) is studied. A comparison of the temperature dependence of the absorbance of free poly(rA) and poly(rA) adsorbed on the nanotube surface [poly(rA)(NT)] at nu(max)= 38,500 cm(-1) shows that the thermostability of the adsorbed polymer is higher. Molecular dynamics simulations demonstrate that more than half of the adenines are not stacked on the tube surface and some of them undergo self-stacking. After addition of a complementary poly(rU) to the poly(rA)(NT) suspension, a double-stranded polymer is formed as confirmed by the characteristic S-like form of its melting curve. However, the melting temperature of this polymer is lower than that of the free poly(rA)poly(rU) duplex. This result indicates that poly(rU) hybridization with poly(rA)(NT) occurs with defects along the whole length of the polymer because of pi-pi stacking between nitrogen bases and the nanotube surface, which hinders the usual hybridization process. Computer modeling demonstrates different possible structures of hybridized polymers on the nanotube surface.

Studies of formation of bivalent copper complexes with native and denatured DNA

The formation of Cu2+ complexes with native and denatured DNA is studied by the methods of differential UV spectroscopy, CD spectroscopy, and viscometry. On ion binding to the bases of native DNA the latter transforms into a new conformation. This transition is accompanied with a sharp increase in UV absorption and a decrease in the intrinsic viscosity though the high degree of helicity persists. Possible sites of Cu2+ ion binding on DNA of various conformations are found along with corresponding constants of complex formation.

Studies of bivalent copper ion binding to poly C

Ultraviolet differential spectra of single-stranded poly C, taken in the presence of Cu2+ ions, are studied at various ionic strengths and temperatures. Coordinational and conformational components of these spectra are obtained. The Cu2+ ion coordination site on the polynucleotide bases is found to be N(3) and possibly O(2). The direction of the poly C absorption band shift due to ion binding and conformational transitions is established. At low ionic strengths of the solution Cu2+ ions cause the helical parts of poly C to melt. At high ones the formation of double-stranded parts was observed in addition to the above effect. The calculated concentration dependences of ion-poly C bases association constants show that binding is cooperative at any ionic strength.

The nature of different influence of Cd2+ ions on the conformational equilibrium of triple-stranded polyribonucleotides poly(U)•poly(A)•poly(U) and poly(I)•poly(A)•poly(I) in aqueous solutions

The influence of Cd2+ ions on the conformational equilibrium of single-stranded (poly(U), poly(A), poly(I)) and triple-stranded polyribonucleotides (A2I, A2U) in aqueous solutions (0.1 M Na+ pH 7) has been investigated using difference UV spectroscopy and thermal denaturation. Analysis of the shape and intensity of the DUV spectra of poly(A), poly(I), and A2I has revealed the presence of two types of complex formed as a result of (i) interaction between Cd2+ and the N7 atoms of purines, producing macrochelates; and (ii) binding of Cd2+ to the N1 atoms of poly(A) and poly(I). Since Cd2+ ions are not bound to heteroatoms of the bases in A2U, the conformation of the structure remains stable up to 0.02 M Cd2+. There is a critical Cd2+ concentration (∼1.5•10−4 M) above which A2I assumes a new helical conformation with lower thermal stability. It is supposed that, upon the formation of the “metallized” A2I triplex, the Cd2+ ions are located inside the triple helix and form bridges between the hypoxanthine and adenine of the homopolynucleotide strands.

Studies of calcium ion binding to poly A

Ultraviolet differential spectra of poly A we studied in the presence of Ca2+ ions with 10(-3)M Na+ in the solution. At concentrations lower than 10(-3)M Ca2+, the ions bind to phosphate groups of the single helical polymer, thus increasing its degree of helicity. At higher concentrations, the ions start binding to the bases of poly A, producing aggregates whose effective radius, as found with an electric microscope, is not more than 10(2) A. These particles stack to form aggregates of an order-of-magnitude higher size. The mutual orientation of bases in the poly A aggregates is of a high degree of order. The calculation of concentration dependences of Ca2+-poly A binding constants shows that this process is cooperative.

DNA conformational equilibrium in the presence of Zn2+ ions in neutral and alkaline solutions.

Effect of Zn(2+) ions on DNA transition from B-form to a metallized form (m-DNA) in Tris and tetraborate buffers at pH 8.5 has been studied by visible and differential UV-spectroscopy and by thermal denaturation. The results have been compared to those obtained at pH 6.5 in cacodylate buffer. It was found that in alkaline solutions Zn(2+) ions induced a hypochromicity of the DNA absorption in the whole spectral range monitored, which was attributed to DNA transition from B- to the m-form. Complete metallization occurred only upon heating the DNA solutions containing more than ~2×10(-4) M of Zn(2+) ions. Phase diagrams of the DNA-zinc complexes at pH 6.5 and 8.5 have been obtained for the first time. The m-DNA form showed higher thermal stability compared to B-DNA.

Divalent metal ion effect on helix-coil transition of high molecular weight DNA in neutral and alkaline solutions

Effect of Mg(2+), Ca(2+), Ni(2+) and Cd(2+) ions on parameters of DNA helix-coil transition in sodium cacodylate (pH 6.5), Tris (pH 8.5) and sodium tetraborate (pH 9.0) buffers have been studied by differential UV-visible spectroscopy and by thermal denaturation. Anomalous behavior of the melting temperature T(m) and the melting interval ΔT in the presence of MgCl(2) was observed in Tris, but not in cacodylate or tetraborate buffers. Changes in the buffer type and pH did not influence T(m) and ΔT dependence on Ca(2+) and Cd(2+) concentrations. Decrease of the T(m) and ΔT of DNA in the presence of Ni(2+) and Cd(2+) was caused by preferential ion interaction with N7 of guanine. This type of interaction was also found for Mg(2+) in Tris buffer. The anomalous decrease in the T(m) and ΔT values was connected to formation of complexes between metal ions and Tris molecules. Transition of DNA single-stranded regions into a compact form with the effective radius of the particles of 300±100 Å was induced by Mg(2+) ions in Tris buffer.

Specific features of Zn2+, Co2+ and Ni2+ ion binding to DNA in alkaline solutions

Dependence of DNA metallization degree during B-DNA transition into the metallized (m) form on DNA concentration has been studied by visible and differential UV-spectroscopy in the presence of Zn(2+), Co(2+) and Ni(2+) ions in tetraborate buffer (pH 8.5) with and without ethidium bromide. Constants of Mt(2+) binding to double stranded DNA were calculated. The obtained binding constants corresponded to the formation of inter-strand metal bridges stabilizing m-form. Thermodynamic origin of higher efficiency of Zn(2+) ions in DNA metallization compared to Co(2+) and Ni(2+) was revealed. Increase of the DNA helix-coil transition temperature by up to 10°С upon formation of m-form in the presence of Zn(2+) ions was observed and rationalized. Furthermore, a strong cooperative decrease (up to 30°С) of the temperature of В→m transition induced by heating in the presence of Zn(2+) was found and its nature was explained.

Interaction of divalent metal ions with poly(riboinosinic acid)

Methods of differential UV and visible spectroscopy are used to study the interaction of Mg 2+ , Ca 2+ , Ma 2+ and Cu 2+ ions with four (4 × 1), and single-stranded poly 1 is solutions with 1 mol.l -1 Na + . Up to concentrations of about 0.1 mol.l -1 Mt 2+ , Mg 3+ and Ca 2+ ions do not bind to hetereatoms of hypoxanthine of 4 × 1, Cu 5+ ions interact with N7 and/or N1 and possibly with O6 through the water molecule of the hydrate shell of the ion. It is likely that the interaction with O6 causes enolization of the hypoxanthine. N1 deprotonation and as a result, this leads to the melting of the four-stranded helix. In single-stranded chains, Cu 2+ ions induce the formation of compact particles which have the effective radii r e ≃ 100 A. The Ma 2+ -induced differential spectra are simalar to those observed in the presence of Cu 2+ ions but in the case of Mn 2+ they occur at concentrations about two orders of magnitude higher. In contrast to the positive cooperativity of the Cu 2+ binding to bases of single-stranded poly I, their interaction with 5 × 1 has the negative cooperativity.

Metallization of single-stranded polyI by Zn(2+) ions in neutral solutions.

Metallization of single-stranded polyinosinic acid (polyI) by Zn(2+) ions at pH 7.0 was studied by differential UV spectroscopy at different temperatures. It was found that polyI is metallized at N7 and N1 atoms of hypoxanthine. The concentration dependence of the degree of binding of Zn(2+) ions to both N7 and N1 sites was obtained, and the corresponding binding constants were determined. Metallization of N1 occurs due to Zn(2+) substituting the imino protons and is effective not only at alkaline but also at neutral pH. This makes multistranded polyI-based systems more promising candidates for use in nanoelectronics than natural DNA sequences, metallization of which can be achieved only at alkaline pH.