Abstract
The binding of three ethidium derivatives, ethidium (1), des-3-amino ethidium (2) and des-8-amino ethidium (3), to short (approximately 35 base pairs), random sequence DNA has been investigated using 300 MHz proton NMR. At 35 degrees C all three drugs cause upfield shifts of the resonances from the exchangeable imino protons, as expected for intercalative binding to DNA. However, the lineshapes vary significantly with the nature of the drug. The temperature dependence of the spectra of the DNA shows that differences between spectra observed at 35 degrees C with ethidium and with des-3-amino ethidium are primarily due to differences in the drug binding kinetics rather than to differences in mode of binding. Removal of the amino group at position 3, but not at position 8, on the parent ethidium shortens the lifetime of the intercalative state; this implies that the 3-NH2 group is involved in stabilization of the drug-DNA complex. Analysis of the drug-DNA spectra indicates that there is a preference for binding of the drugs adjacent to G.C base pairs.
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Selected References
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- Aktipis S., Kindelis A. Optical properties of the deoxyribonucleic acid-ethidium bromide complex. Effect of salt. Biochemistry. 1973 Mar 13;12(6):1213–1221. doi: 10.1021/bi00730a031. [DOI] [PubMed] [Google Scholar]
- Baguley B. C., Falkenhaug E. M. The interaction of ethidium with synthetic double-stranded polynucleotides at low ionic strength. Nucleic Acids Res. 1978 Jan;5(1):161–171. doi: 10.1093/nar/5.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bresloff J. L., Crothers D. M. DNA-ethidium reaction kinetics: demonstration of direct ligand transfer between DNA binding sites. J Mol Biol. 1975 Jun 15;95(1):103–123. doi: 10.1016/0022-2836(75)90339-3. [DOI] [PubMed] [Google Scholar]
- Early T. A., Kearns D. R. 1H nuclear magnetic resonance investigation of flexibility in DNA. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4165–4169. doi: 10.1073/pnas.76.9.4165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eron L. J., McAuslan B. R. Inhibition of deoxyribonuclease action by actinomycin D and ethidium bromide. Biochim Biophys Acta. 1966 Mar 21;114(3):633–636. doi: 10.1016/0005-2787(66)90113-4. [DOI] [PubMed] [Google Scholar]
- Giessner-Prettre C., Pullman B. Intermolecular nuclear shielding values for protons of purines and flavins. J Theor Biol. 1970 Apr;27(1):87–95. doi: 10.1016/0022-5193(70)90130-x. [DOI] [PubMed] [Google Scholar]
- Hogan M. E., Jardetzky O. Effect of ethidium bromide on deoxyribonucleic acid internal motions. Biochemistry. 1980 May 13;19(10):2079–2085. doi: 10.1021/bi00551a012. [DOI] [PubMed] [Google Scholar]
- Jain S. C., Tsai C. C., Sobell H. M. Visualization of drug-nucleic acid interactions at atomic resolution. II. Structure of an ethidium/dinucleoside monophosphate crystalline complex, ethidium:5-iodocytidylyl (3'-5') guanosine. J Mol Biol. 1977 Aug 15;114(3):317–331. doi: 10.1016/0022-2836(77)90253-4. [DOI] [PubMed] [Google Scholar]
- Jones R. L., Lanier A. C., Keel R. A., Wilson W. D. The effect of ionic strength on DNA-ligand unwinding angles for acridine and quinoline derivatives. Nucleic Acids Res. 1980 Apr 11;8(7):1613–1624. doi: 10.1093/nar/8.7.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kastrup R. V., Young M. A., Krugh T. R. Ethidium bromide complexes with self-complementary deoxytetranucleotides. Demonstration and discussion of sequence preferences in the intercalative binding of ethidium bromide. Biochemistry. 1978 Nov 14;17(23):4855–4865. doi: 10.1021/bi00616a002. [DOI] [PubMed] [Google Scholar]
- Kearns D. R. High-resolution nuclear magnetic resonance studies of double helical polynucleotides. Annu Rev Biophys Bioeng. 1977;6:477–523. doi: 10.1146/annurev.bb.06.060177.002401. [DOI] [PubMed] [Google Scholar]
- Krugh T. R., Reinhardt C. G. Evidence for sequence preferences in the intercalative binding of ethidium bromide to dinucleoside monophosphates. J Mol Biol. 1975 Sep 15;97(2):133–162. doi: 10.1016/s0022-2836(75)80031-3. [DOI] [PubMed] [Google Scholar]
- LERMAN L. S. Structural considerations in the interaction of DNA and acridines. J Mol Biol. 1961 Feb;3:18–30. doi: 10.1016/s0022-2836(61)80004-1. [DOI] [PubMed] [Google Scholar]
- Müller W., Crothers D. M. Interactions of heteroaromatic compounds with nucleic acids. 1. The influence of heteroatoms and polarizability on the base specificity of intercalating ligands. Eur J Biochem. 1975 May;54(1):267–277. doi: 10.1111/j.1432-1033.1975.tb04137.x. [DOI] [PubMed] [Google Scholar]
- Olmsted J., 3rd, Kearns D. R. Mechanism of ethidium bromide fluorescence enhancement on binding to nucleic acids. Biochemistry. 1977 Aug 9;16(16):3647–3654. doi: 10.1021/bi00635a022. [DOI] [PubMed] [Google Scholar]
- Patel D. J., Canuel L. L. Biphasic helix-coil transition of the ethidium bromide-poly (dA-dT) and the propidium diiodide - poly (dA-dT) complexes. Stabilization of base-pair regions centered about the intercalation site. Biopolymers. 1977 Apr;16(4):857–873. doi: 10.1002/bip.1977.360160410. [DOI] [PubMed] [Google Scholar]
- Patel D. J., Canuel L. L. Ethidium bromide-(dC-dG-dC-dG)2 complex in solution: intercalation and sequence specificity of drug binding at the tetranucleotide duplex level. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3343–3347. doi: 10.1073/pnas.73.10.3343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reinhardt C. G., Krugh T. R. A comparative study of ethidium bromide complexes with dinucleotides and DNA: direct evidence for intercalation and nucleic acid sequence preferences. Biochemistry. 1978 Nov 14;17(23):4845–4854. doi: 10.1021/bi00616a001. [DOI] [PubMed] [Google Scholar]
- Tsai C. C., Jain S. C., Sobell H. M. Visualization of drug-nucleic acid interactions at atomic resolution. I. Structure of an ethidium/dinucleoside monophosphate crystalline complex, ethidium:5-iodouridylyl (3'-5') adenosine. J Mol Biol. 1977 Aug 15;114(3):301–315. doi: 10.1016/0022-2836(77)90252-2. [DOI] [PubMed] [Google Scholar]
- Wakelin L. P., Waring M. J. Kinetics of drug-DNA interaction. Dependence of the binding mechanism on structure of the ligand. J Mol Biol. 1980 Dec 5;144(2):183–214. doi: 10.1016/0022-2836(80)90032-7. [DOI] [PubMed] [Google Scholar]
- Wang J. C. The degree of unwinding of the DNA helix by ethidium. I. Titration of twisted PM2 DNA molecules in alkaline cesium chloride density gradients. J Mol Biol. 1974 Nov 15;89(4):783–801. doi: 10.1016/0022-2836(74)90053-9. [DOI] [PubMed] [Google Scholar]
- Waring M. J. Complex formation between ethidium bromide and nucleic acids. J Mol Biol. 1965 Aug;13(1):269–282. doi: 10.1016/s0022-2836(65)80096-1. [DOI] [PubMed] [Google Scholar]
