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Related Experiment Videos

Nucleotide conformational change induced by cationic bilayers.

Iseli L Nantes1, Felipe M Correia, Adelaide Faljoni-Alario

  • 1Centro Interdisciplinar de Investigação Bioqui;mica, CIIB, Universidade de Mogi das Cruzes, SP, Mogi das Cruzes, Brazil.

Archives of Biochemistry and Biophysics
|July 16, 2003
PubMed
Summary
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Cationic lipids like DODAB bilayers alter the conformation of 2'-deoxyadenosine 5'-monophosphate (dAMP) from an anti to a syn form. This change, observed via CD spectroscopy and molecular modeling, may explain DNA denaturation when complexed with cationic lipids.

Area of Science:

  • Biophysical Chemistry
  • Molecular Biophysics
  • Supramolecular Chemistry

Background:

  • Understanding the molecular interactions between nucleic acids and cationic lipids is crucial for developing gene delivery systems.
  • The conformation of nucleotides influences DNA structure and function, including processes like denaturation.
  • Cationic lipids form aggregates like micelles and bilayers that can interact with charged molecules such as nucleotides.

Purpose of the Study:

  • To investigate the impact of cetyltrimethylammonium bromide (CTAB) micelles and dioctadecyldimethylammonium bromide (DODAB) bilayers on the molecular conformation of 2 -deoxyadenosine 5 -monophosphate (dAMP).
  • To elucidate the role of medium polarity and phosphate charge on dAMP conformation.
  • To explore the potential link between dAMP conformational changes and DNA denaturation induced by cationic lipids.

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Main Methods:

  • Circular dichroism (CD) spectroscopy was employed to analyze dAMP conformation under various conditions.
  • Molecular modeling was used to simulate dAMP conformations with minimal energy by varying torsion angles.
  • Experiments and modeling considered different dielectric constants (E) and phosphate charges (C) of the medium (E80_C2, E80_C0, E1_C2, E1_C0).

Main Results:

  • Decreased medium polarity led to reduced intensity of the negative band in dAMP CD spectra (190-210 nm).
  • Increased dAMP:DODAB ratio enhanced the positive band (210-230 nm) and caused a red shift, indicating increased base stacking similar to poly(A).
  • DODAB bilayers induced base stacking and insertion, forcing dAMP from an extended anti conformation to a syn conformation, unlike CTAB micelles. Modeling confirmed this anti-to-syn transition.

Conclusions:

  • Cationic DODAB bilayers induce a significant conformational change in dAMP from anti to syn, driven by base stacking and insertion into the lipid aggregate.
  • This dAMP conformational shift, observed through CD spectroscopy and molecular modeling, is a key finding.
  • The study suggests that this nucleotide conformational change may be the underlying mechanism for DNA denaturation observed upon complexation with cationic lipids.