Abstract
Chemical and thermal denaturation of calf thymus DNA (as a multidomain macromolecule) have been investigated in the presence of high concentrations of dimethyltin dichloride (Me2SnCl2) over the temperature range (55–95°C) in 50.0 mM phosphate buffer at pH 7.6 using temperature scanning spectroscopy and calorimetry methods. Results showed that over the concentration range of 6–16 mM, Me2SnCl2 is a chemical denaturant and denatures the double-strand DNA in a three-state manner. The denaturation data are analyzed based on the effective Gibbs free energy (ΔG°eff) approach and the chemical denaturation parameters including ΔG°eff, m value and equilibrium unfolding constant (K U) were obtained. Ultraviolet (UV) melting curves of the DNA at 260 nm as well as the calorimetric measurements were used to estimate the binding constants (K), melting enthalpy (ΔH°m) and binding enthalpy (ΔH°b). Furthermore, at low concentrations (up to 5 mM), Me2SnCl2 binds to the phosphate groups of DNA in an exothermic step and had no significant effect on double-strand DNA stability, confirmed by the fact that the T m value did not change. However, high (denaturing) concentrations of Me2SnCl2 (more than 9 mM) caused considerable destabilization of DNA associated with the formation of a partially unfolded intermediate at 13.6 mM of Me2SnCl2. The formed intermediate showed a lower thermal transition temperature (T m) by a magnitude of 10°C in relation to the native DNA. Finally, a new correlation is introduced for interpretation of thermal denaturation behavior of calf thymus DNA over the whole range of ligand (Me2SnCl2) concentration (0–16 mM).
Similar content being viewed by others
References
Ahmad F, Salahuddin A (1976) Reversible unfolding of the major fraction of ovalbumin by guanidine hydrochloride. Biochemistry 15:5168
Arscott PG, Li AZ, Bloomfield VA (1990) Condensation of DNA by trivalent cations, 1: Effects of DNA length and topology on the size and shape of condensed particles. Biopolymers 30:619–630
Backmann J, Schafer G, Wyns L, Bonisch H (1998) The stability of salt bridges at high temperatures: implications for hyperthermophilic proteins. J Mol Biol 284:817–833
Barbieri R, Sikvestri A (1991) The interaction of native DNA with dimethyltin (IV) species. J Inorg Biochem, 41:31–35
Barbieri R, Alonzo G, Herber RH (1987) The configuration and lattice dynamics of complexes of dialkyltin(IV) with adenosine 5′-monophosphate and phenyl phosphates. J Chem Soc Dalton Trans 789–794
Barbieri R, Silvestri A, Piro V, (1990) Tin 119Sn: Mossbauer titration of dimethyltin-and trimethyltin (IV) hydroxides with model ligands mimicking nucleic acid phosphate sites, and with deoxyribonucleic acid. J Chem Soc Dalton Trans 3605–3609
Barbieri R, Silvestri A, Giuliani AM, Piro V (1992) Organotin compounds and deoxyribonucleic acid. V J Chem Soc Dalton Trans 585–590
Barone G, Barbieri R, LaManna G, Kock MHJ (2000) The interaction of deoxyribonucleic acid with methyltin (IV) moieties in solution studied by small-angle X-ray scattering, circular dichroism and UV spectroscopy. Appl Organometal Chem 14:189–196
Basu HS, Pellarin M, Feuerstein BG, Shirahata A, Samejima K, Deen DF, Martin LJ (1993) Interaction of a polyamine analogue, 1,19-bis-(ethylamino)-5,10,15-triazanonadecane (BE-4-4-4-4), with DNA and effect on growth, survival, and polyamine levels in seven human brain tumor cell lines. Cancer Res 53:3948–3955
Bathaie SZ, Moosavi-Movahedi AA, Ranjbar B, Saboury AA (2003) A mechanistic study of the histone H1-DNA complex dissociation by sodium dodecyl sulfate. Colloids Surfaces B 28:17–25
Bathaie SZ, Moosavi-Movahedi AA, Saboury AA (1999) Energetic and binding properties of DNA upon interaction with dodecyl trimethylammonium bromide. Nucleic Acids Res 27:1001–1005
Bhattacharya S, Mandal SS (1997) Interaction of surfactants with DNA. Role of hydrophobicity and surface charge on intercalation and DNA melting. Biochim Biophys Acta 1323:29–44
Blake RD, Delcourt SG (1998) Thermal stability of DNA. Nucleic Acids Res 26(14):3323–3332
Blunden SJ, Chapman A (1986) Organometallic Compounds in the Environment (Ed PJ Craig). Wiley, New York, Ch 3, pp 111–159
Blunden SJ, Cusack PA, Hill R (1985) The Industrial Uses of Tin Chemicals, the Royal Society of Chemistry, London
Boudker O, Todd MJ, Freire E (1997) The structural stability of the co-chaperonin GroES, J Mol Biol 272:770–779
Carra JH, Privalov PL (1995) Energetics of denaturation and m values of staphylococcal nuclease mutants. Biochemistry 34:2034–2041
Casini A, Messori L, Orioli P, Gielen M, Kemmer M, Willem R (2001) Interactions of two cytotoxic organotin (IV) compounds with calf thymus DNA. J Inorg Biochem, 85(4):297–300
Clarke MJ, Zhu F, Frasca DR (1999) Non-platinum chemotherapeutic metallopharmaceuticals. Chem Rev 99:2511–2533
Duguid JG, Bloomfield VA, Benevides JM, Thomas Jr. GJ (1995) Raman spectroscopy of DNA-metal complexes, II: The thermal denaturation of DNA in the presence of Sr2+, Ba2+, Mg2+, Ca2+, Mn2+, Co2+, Ni2+ and Cd2+. Biophys J 69:2623–2641
Frank-Kamenetskii F (1971) Simplification of the empirical relationship between melting temperature of DNA, its GC content and concentration of sodium ions in solution. Biopolymers 10(12):2623–2624
Freifelder D (1986) Molecular Biology. Jones and Bartlett, USA
Ghaemmaghami S, Fitzgerald MC, Oas TG (2000) A quantitative, high-throughput screen for protein stability. Proc Natl Acad Sci 97:8296–8301
Greene RFJ, Pace CN (1974) Urea and guanidine hydrochloride denaturation of ribonuclease, lysozyme, [alpha]-chymotrypsin, and [beta]-lactoglobulin. J Biol Chem 249:5388–5393
Gruenwedel DW (1974) Salt effects on the denaturation of DNA: A calorimetric investigation of the transition enthalpy of calf thymus DNA in Na2SO4 solutions of varying ionic strength. Biochim Biophys Acta 340(1):16–30
Guthe S, Kapinos L, Moglish A, Meier S, Grzesiek S, Kiefhaber T (2004) Very fast folding and association of a trimerization domain from bacteriophage T4 fibritin, J Mol Biol 337:905–915
Hillen W, Goodman TC, Wells RD (1981) Salt dependence and thermodynamic interpretation of the thermal denaturation of small DNA restriction fragments. Nucleic Acids Res 9(2):415–436
Hofr C, Brabec V (2001) Thermal and thermodynamic properties of duplex DNA containing site-specific interstrand cross-link of antitumor cisplatin or its clinically ineffective trans isomer. J Biol Chem 276:9655–9661
Hynes MJ, Dowd MO (1987) Interactions of the trimethyltin (IV) cation with carboxylic acids, amino acids, and related ligands. J Chem Soc Dalton Trans 563–566
Inman RB, Baldwin RL (1964) Helix-random coil transitions in DNA homopolymer pairs. J Mol Biol 8:452–469
Jenkins SM, Ehman K, Barone S (2004) Structure–activity comparison of organotin species: dibutyltin is a developmental neurotoxicant in vitro and in vivo. Develop Brain Res 151(1–2):1–12
Johnson CR, Morin PE, Arrowsmith CH, Freire E (1995) Thermodynamic analysis of the structural stability of the tetrameric oligomerization domain of p53 tumor suppressor. Biochemistry 34:5309–5316
Korolev N, Lyubartsev AP, Nordenskio L (1998) Application of polyelectrolyte theories for analysis of DNA melting in the presence of Na+ and Mg2+ ions. Biophys J 75:3041–3056
Korolev N, Vlasov AP, Kuznetsov IA (1994) Thermal denaturation of Na- and Li-DNA in salt free solutions. Biopolymers 34:1275–1290
Leng F, Chaires JB, Waring MJ (2003) Energetics of echinomycin binding to DNA. Nucleic Acids Res 31(21):6191–6197
Leng F, Priebe W, Chaires JB (1998) Ultratight DNA binding of a new bisintercalating anthracycline antibiotic. Biochemistry 37:1743–1753
Lobo BA, Davis A, Koe G, Smith JG, Middaugh CR (2001) Isothermal titration calorimetric analysis of the interaction between cationic lipids and plasmid DNA, Arch Biochem Biophys 386:95–105
Lovett CM Jr, Fitzgibbon TN, Chang R (1989) Effect of UV irradiation on DNA as studied by its thermal denaturation. J Chem Edu 66(6)
Luck G, Zimmer C (1972) Conformational aspects and reactivity of DNA: Effects of manganese and magnesium ions on interaction with DNA. Eur J Biochem 29:528–536
Mahmoudi A, Nazari K, Mohammadian N, Safarian S, Moosavi-Movahedi AA (2000) The relationship of function and structural stability of horseradish peroxidase by interaction with Cu2+ and Ni2+ ions. Biophys J 78(1):1748
Makhatadze GI, Privalov PL (1993) Contribution of hydration to protein folding thermodynamics, I: The enthalpy of hydration. J Mol Biol 232:639–659
Marmur J, Doty P (1962) Determination of the base composition of deoxyribonucleic acid from its thermal denaturation temperature. J Mol Biol 5:109–118
Mascotti DP, Lohman TM (1997) Thermodynamics of oligoarginines binding to RNA and DNA. Biochemistry 36:7272–7279
McFaydyen WD, Sotirellis N, Denny WA, Wakelin PG (1990) The interaction of substituted and rigidly linked diquinolines with DNA. Biochim Biophys Acta 1048:50–58
McGhee JD (1976) Theoretical calculations of the helix-coil transition of DNA in the presence of large, cooperatively binding ligands. Biopolymers 15:1345–1375
Mildvan AS, Loeb LA (1979) The role of metal ions in the mechanisms of DNA and RNA polymerases. CRC Crit Rev Biochem 6:219–244
Moosavi-Movahedi AA, Golchin AR, Nazari K, Saboury AA (2004) Chamani, Microcalorimetry, energetics and binding studies of DNA-dimethyltin dichloride complexes, J and Tangestani-Nejad, S. Thermochimica Acta 414:233–241
Nazari K, Moosavi-Movahedi AA (2000) Potentiometric titration and enthalpy evaluation of horseradish peroxidase in the presence of n-dodecyl trimethylammonium bromide. Colloids Surfaces B 18:63–70
Nazari K, Golchin AR, Moosavi-Movahedi AA, Saboury AA, Shokravi A, Tangestani-Nejad (2005) Microcalorimetry and binding studies of DNA upon interaction with [pyridine diamine] 2[co(phenanthroline dicarboxylate)2]. Thermochimica Acta 220(3):292–299
Pace CN (1990) Measuring and increasing protein stability. Trends Biotech 8:93
Pace NC (1986) Determination and analysis of urea and guanidine hydrochloride denaturation curves. Methods Enzymol 131:266–280
Panse VG, Swaminathan CP, Aloor JJ, Surolia A, Varadarajan R (2000) Unfolding thermodynamics of the tetrameric chaperone SecB. Biochemistry 39:2362–2369
Park C, Marqusee S (2004) Analysis of the stability of multimeric proteins by effective ΔG and effective m-values. Prot Sci 13:2553–2558
Pellerito C, Nagy L, Pellerito L, Szorcsik A (2006) Biological activity studies on organotin (IV)n+ complexes and parent compounds. J Organomet Chem 691(8):1733–1747
Penninks AH, Seinen W (1984) Mechanisms of dialkyltin induced immunopathology, Vet Q 6:209–219
Pezzano H, Pato F (1980) Structure of binary complexes of mono- and polynucleotides with metal ions of the first transition group. Chem Rev 80:365–401
Privalov PL (1979) Stability of proteins: small globular proteins. Adv Prot Chem 33:167–197
Ptitsyn OB (1992) The molten globule state in Protein Folding, (Ed. Creighton TE), Freeman, New York, pp 243–300
Ptitsyn OB (1995) Molten globule and protein folding. Adv Protein Chem 47:83–229
Ragone R (2000) How the protein concentration affects unfolding curves of oligomers. Biopolymers 53:221–225
RamakarishnanV (1997) Histone structure and the organization of the nucleosome. Annu Rev Biophys Biomol Struct 26:83–112
Raspud E, Olvera M, Cruz de la Silkarav JL, Livolan F (1998) Precipitation of DNA by polyamines: A polyelectrolyte behavior. Biophys J 74:381–393
Santoro MM, Bolen DW (1992) A test of the linear extrapolation of unfolding free energy changes over an extended denaturant concentration range. Biochemistry 31:4901–4907
Schellman J (1978) Solvent denaturation. Biopolymers 17:1305–1322
Schildkraut C (1965) Dependence of the melting temperature of DNA on salt concentration. Biopolymers 3:195–208
Schultze P, Hud NV, Smith FW, Feigon J (1999) The effect of sodium, potassium and ammonium ions on the conformation of the dimeric quadruplex formed by the Oxytricha nova telomere repeat oligonucleotide d(G(4)T(4)G(4)). Nucleic Acids Res 27(15):3018–3028
Shchyolkina AK, Borisova OF, Livshits MA, Pozmogova GE, Chernov BK, Klement R, Jovin TM (2000) Parallel-stranded DNA with mixed AT/GC composition: role of trans G·C base pairs in sequence dependent helical stability. Biochemistry 39:10034–10044
Shortle D (1989) Probing the determinants of protein folding and stability with amino acid substitutions. J Biol Chem 264:5315–5318
Silinski P, Allingham MG, Fitzgerald MC (2001) Guanidine-induced equilibrium unfolding of a hexameric enzyme 4-oxalocrotonate tautomerase (4-OT). Biochemistry 40:4493–4502
Soulages JL (1998) Chemical denaturation: Potential impact of undetected intermediates in the free energy of unfolding and m-values obtained from a two-state assumption. Biophys J 75:484–492
Syng-ai C, Basu Baul TS, Chatterjee A (2001) Inhibition of cell proliferation and antitumor activity of a novel organotin compound, J Environ Pathol Toxicol Oncol 20(4):333–342
Tabassum S, Pettinari C (2006) Chemical and biotechnological developments in organotin cancer chemotherapy. J Organomet Chem 691(8):1761–1766
Yao M, Bolen DW (1995) How valid are denaturant-induced unfolding free energy measurements? Level of conformance to common assumptions over an extended range of ribonuclease A stability. Biochemistry 34:3771–3781
Acknowledgements
The financial support of the research council of Islamic Azad University, the research council of Tehran University and the Iran National Science Foundation are gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nazari, K., Gholami, N. & Moosavi-Movahedi, A.A. Stability of DNA upon interaction with dimethyltin dichloride. Med Chem Res 16, 238–257 (2007). https://doi.org/10.1007/s00044-007-9028-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00044-007-9028-5