Dynamic light scattering and fluorescence study of the interaction between double-stranded DNA and poly(amido amine) dendrimers.
The interaction between a cationic poly(amido amine) (PAMAM) dendrimer of generation 4 and double-stranded salmon sperm DNA in 10 mM NaBr solution has been investigated using dynamic light scattering (DLS) and steady-state fluorescence spectroscopy. The structural parameters of the formed aggregates as well as the complex formation process were studied in dilute solutions. When DNA is mixed with PAMAM dendrimers, it undergoes a transition from a semiflexible coil to a more compact conformation due to the electrostatic interaction present between the cationic dendrimer and the anionic polyelectrolyte. The DLS results reveal that one salmon sperm DNA molecule forms a discrete aggregate in dilute solution with several PAMAM dendrimers with a mean apparent hydrodynamic radius of 50 nm. These discrete complexes coexist with free DNA at low molar ratios of dendrimer to DNA, which shows that cooperativity is present in the complex formation. The formation of the complexes was confirmed by agarose gel electrophoresis measurements. DNA in the complexes was also found to be significantly more protected against DNase catalyzed digestion compared to free DNA. The number of dendrimers per DNA chain in the complexes was found to be approximately 35 as determined by steady-state fluorescence spectroscopy.
Orberg
Marie-Louise
ML
Physical Chemistry 1, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden. marie-louise.orberg@fkem1.lu.se
Schillén
Karin
K
Nylander
Tommy
T
D000818:Animals; D004247:DNA / Q000737:chemistry*; D050091:Dendrimers; D003851:Deoxyribonucleases / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005453:Fluorescence; D008027:Light; D008297:Male; D011073:Polyamines / Q000737:chemistry*; D012474:Salmon; D012542:Scattering, Radiation; D012996:Solutions / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013094:Spermatozoa / Q000737:chemistry
D050091:Dendrimers; C104700:PAMAM Starburst; D011073:Polyamines; D012996:Solutions; D004247:DNA; D003851:Deoxyribonucleases