NMR investigation of the complexation of daunomycin with deoxytetranucleotides of different base sequence in aqueous solution.
500 MHz NMR spectroscopy has been used to investigate the complexation of the anthracycline antibiotic daunomycin (DAU) with self-complementary deoxytetranucleotides, 5'-d(CGCG), 5'-d(GCGC), 5'-d(TGCA), 5'-d(ACGT) and 5'-d(AGCT), of different base sequence in aqueous salt solution. 2D homonuclear 1H NMR spectroscopy (TOCSY and NOESY) and heteronuclear 1H - 31P NMR spectroscopy (HMBC) have been used for complete assignment of the non-exchangeable protons and the phosphorus resonance signals, respectively, and for a qualitative determination of the preferred binding sites of the drug. Analysis shows that DAU intercalates preferentially into the terminal sites of each of the tetranucleotides and that the aminosugar of the antibiotic is situated in the minor groove of the tetramer duplex, partly eclipsing the third base pair. A quantitative determination of the complexation of DAU with the deoxytetranucleotides has been made using the experimental concentration and temperature dependences of the drug proton chemical shifts; these have been analysed in terms of the equilibrium reaction constants, limiting proton chemical shifts and thermodynamical parameters (enthalpies deltaH, entropies deltaS) of different drug-DNA complexes (1:1, 1:2, 2:1, 2:2) in aqueous solution. It is found that DAU interacts with sites containing three adjacent base pairs but does not show any significant sequence specificity of binding with either single or double-stranded tetranucleotides, in contrast with other intercalating drugs such as proflavine, ethidium bromide and actinomycin D. The most favourable structures of the 1:2 complexes have been derived from the induced limiting proton chemical shifts of the drug in the intercalated complexes with the tetranucleotide duplex, in conjunction with 2D NOE data. It has been found that the conformational parameters of the double helix and the orientation of the DAU chromophore in the intercalated complexes depend on base sequence at the binding site of the tetramer duplexes in aqueous solution.
Davies
D B
DB
School of Biological and Chemical Sciences, Birkbeck College, University of London, UK. a.king@chem.bsk.ac.uk
Eaton
R J
RJ
Baranovsky
S F
SF
Veselkov
A N
AN
D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D003630:Daunorubicin / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008962:Models, Theoretical; D009711:Nucleotides / Q000378:metabolism*; D013696:Temperature; D013816:Thermodynamics; D014867:Water / Q000378:metabolism*
D000903:Antibiotics, Antineoplastic; D009711:Nucleotides; D014867:Water; D004247:DNA; D003630:Daunorubicin