Dynamics of peptide - nucleic acid complexes. Fluorescence polarization studies.
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Chemical Phenomena↗
Biomedical subjects
J C Brochon
Publications and source records attributed to J C Brochon.
[Interactions of HIV-1 DNA heterocyclic bases with viral DNA].
Human immunodeficiency virus type 1 integrase is one of three viral enzymes, and it realizes a key process of the viral replication cycle, i.e. viral DNA integration into infected cell genome. Integrase recognizes nucleotide sequences located at the ends of the viral DNA U3 and U5 LTRs and catalyzes 3'-processing and strand transfer reactions. To study the interactions between integrase and viral DNA at present work, we used modified integrase substrates mimicking the terminal U5 LTR sequence and containing non-nucleoside insertions in one or/and both strands. It is shown that the substrate modifications have no influence on the integrase binding rate, while the heterocyclic bases removal in the 5th and 6th substrate positions and in the 3rd position of the substrate processed strand distinctly inhibits the integrase catalytic activity. This fact demonstrates these bases significance for the active enzyme/substrate complex formation. On the contrary, modification of the 3rd position within substrate non-processed strand stimulates 3'-processing. Since heterocyclic base elimination results in disruption of the DNA complementary and staking interactions, this result shows that DNA double helix destabilization close to the cleaved bond promotes the 3'-processing.
DNA, Viral↗
[Intensity and anisotropy decays measurement of perylene in pigeon erythrocyte membrane. Comparison with an isotropic viscous medium (author's transl)].
Using synchrotron radiation as the excitation light, we studied the fluorescence parameters of perylene incubated with pigeon erythrocyte membranes and with an isotropic viscous medium, the Primol 342 oil. From 4 to 37 degrees C, we observed a single lifetime of 4.5 ns in the oil and two with the membrane (tau 1 = 1-1.4 ns and tau 2 = 5.4-6.1 ns). The dependence upon temperature of the rotation correlation time of perylene (theta) in the oil was characteristic of an isotropic medium, whereas the limiting value of anitropy (r infinity) was zero. With the membrane, r infinity decreased from 0.14 to 0.06 and theta from 2.9 to 0.5 ns, indicating a greater amplitude and frequency of molecular motions. The addition of chlorpromazine, indomethacine, tetracaine, n-octylamine, octanol or octanoic acid to the membrane decreased the tau 1 and tau 2 values. This would stem from the disorganization of the membrane induced by the drugs.
Animals↗