An infrared spectroscopic study of the effect of hydration on cationic lipid/DNA complexes.
Infrared spectroscopy was used to examine the effect of dehydration on the structure of DNA and cationic lipid/DNA complexes (CLDCs). Information regarding the effect of hydration on the interface between the cationic lipids and DNA was obtained by following subtle but reproducible changes in vibrational bands arising from the DNA bases and phosphate backbone as well as bands from the lipid ester groups within the interfacial region of the bilayer. Dehydration of supercoiled plasmid DNA induces a transition from a B-conformation in solution to a mixed conformation in the dried state. Changes in vibrations of the bases upon drying suggest a change to an A-conformation whereas vibrations from the phosphate moieties suggest A- or C-forms. Vibrational changes in the ribose ring suggest adoption of a C-conformation. When CLDCs composed of either DOTAP (1,2-dioleoyl-3-trimethylammonium-propane) or DDAB (dioctadecyldimethylammonium bromide) cationic lipids with or without equimolar amounts of the helper lipids cholesterol or DOPE (1,2-dioleoylphosphatidylethanolamin) are dried, the DNA is still able to undergo these structural transitions suggesting a nonrigid CLDC structure. The effect of dehydration on these interfacial interactions was found to be dependent on the type of cationic lipid used as well as the type of helper lipid. In addition, this work provides a simple spectroscopic analytical approach that can be used for the characterization of nonviral vectors that has potential pharmaceutical utility.
Choosakoonkriang
Sirirat
S
The Department of Pharmaceutical Chemistry, The University of Kansas, 2095 Constant Avenue, Lawrence, Kansas 66047, USA.
Wiethoff
Christopher M
CM
Koe
Gary S
GS
Koe
Janet G
JG
Anchordoquy
Thomas J
TJ
Middaugh
C Russell
CR
D002412:Cations / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002626:Chemistry, Pharmaceutical; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D003890:Desiccation / Q000379:methods; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis* / Q000737:chemistry; D017433:Protein Structure, Secondary; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods
D002412:Cations; D008055:Lipids; D004247:DNA