Quartz crystal microbalance with dissipation monitoring and spectroscopic ellipsometry measurements of the phospholipid bilayer anchoring stability and kinetics of hydrophobically modified DNA oligonucleotides.
Decorating lipid bilayers with oligonucleotides has great potential for both fundamental studies and applications, taking advantage of the membrane properties and the specific Watson-Crick base pairing. Here, we systematically studied the binding of DNA oligonucleotides with the frequently used hydrophobic anchors cholesterol, stearyl, and distearyl to supported lipid bilayers made of dioleoylphosphatidylcholine (DOPC) by quartz crystal microbalance with dissipation monitoring and spectroscopic ellipsometry (SE). All three anchors were found to incorporate well into DOPC lipid membranes, yet only the distearyl-based anchor remained stable in the bilayer when it was rinsed. The unstable anchoring of the cholesterol- and stearyl-based oligonucleotides can, however, be stabilized by hybridization of the oligonucleotides to complementary DNA modified with a second hydrophobic anchor of the same type. In all cases, the incorporation into the lipid bilayer was found to be limited by mass transport, although micelle formation likely reduced the effective concentration of available oligonucleotides in some samples, leading to substantial differences in binding rates. Using a viscoelastic model to determine the thickness of the DNA layer and elucidating the surface coverage by SE, we found that at equal bulk concentrations double-stranded DNA constructs attached to the lipid bilayer establish a layer that is thicker than that of single-stranded oligonucleotides, whereas the DNA surface densities are similar. Shortening the length of the oligonucleotides, on the other hand, does alter both the thickness and surface density of the DNA layer. This indicates that at the bulk oligonucleotide concentrations employed in our experiments, the packing of the oligonucleotides is not affected by the anchor type, but rather by the length of the DNA. The results are useful for material and biomedical applications that require efficient linking of oligonucleotides to lipid membranes.
FOM Institute AMOLF , Science Park 104, 1098 XG Amsterdam, The Netherlands.
Stef A J
SA
van der Meulen
Galina V
GV
Dubacheva
Marileen
M
Dogterom
Ralf P
RP
Richter
Mirjam E
ME
Leunissen
D004247:DNA / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008051:Lipid Bilayers / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D058525:Quartz Crystal Microbalance Techniques / Q000379:methods*
D008051:Lipid Bilayers; D009841:Oligonucleotides; D010713:Phosphatidylcholines; D004247:DNA; C017251:1,2-oleoylphosphatidylcholine