Formation of DNA nanoparticles in the presence of novel polyamine analogues: a 
laser light scattering and atomic force microscopic study

Vijayanathan, V.; Thomas, T.; Antony, T.; Shirahata, A.; Thomas, T. J.

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Formation of DNA nanoparticles in the presence of novel polyamine analogues: a laser light scattering and atomic force microscopic study

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Thomas, T.
Department of Molecular Cell Biology, MPI for biophysical chemistry, Max Planck Society;

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Antony, T.
Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society;

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Vijayanathan, V., Thomas, T., Antony, T., Shirahata, A., & Thomas, T. J. (2004). Formation of DNA nanoparticles in the presence of novel polyamine analogues: a laser light scattering and atomic force microscopic study. Nucleic Acids Research, 32(1): DOI: 10.1093/nar/gkg936, pp. 127-134. Retrieved from http://nar.oxfordjournals.org/cgi/content/full/32/1/127.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0012-ED23-D

Abstract

We synthesized a pentamine (3-3-3-3) and two hexamine (3-3-3-3-3 and 3-4-3-4-3) analogues of the natural polyamine, spermine (3-4-3) and studied their effectiveness in condensing pGL3 plasmid DNA, using light scattering and atomic force microscopic (AFM) techniques. The midpoint concentration of the polyamines on pGL3 condensation (EC50) was 11.3, 10.6, 1.5, 0.49 and 0.52 mM, respectively, for 3-4-3, norspermine (3-3-3), 3-3-3-3, 3-3-3-3-3 and 3-4-3-4-3 in 10 mM Na cacodylate buffer. Dynamic laser light scattering study showed a decrease in hydrodynamic radii of plasmid DNA particles as the number of positive charges on the polyamines increased. AFM data showed the presence of toroids with outer diameter of 117-191 nm for different polyamines, and a mean height of 2.61 ± 0.77 nm. AFM results also revealed the presence of intermediate structures, including those showing circumferential winding of DNA to toroids. The dependence of the EC₅₀ on Na⁺ concentration suggests different modes of binding of spermine and its higher valent analogues with DNA. Our results show a 20-fold increase in the efficacy of hexamines for DNA condensation compared to spermine, and provide new insights into the mechanism(s) of DNA nanoparticle formation. These studies might help to develop novel nonviral gene delivery vehicles.