文摘
Membrane fusion events are essential to cell biology, and a number of reductionist systems have been developed to mimic the behavior of these biological motifs. One such system monitors the DNA hybridization-mediated fusion of liposomes with the liquid crystal (LC) interface by observing changes in LC orientation using a simple optical detection scheme. We have systematically explored key parameters of this system to determine their effects on individual elementary steps of the complex fusion mechanism. The liposome composition, specifically the degree of lipid unsaturation and PE content, decreased the bilayer rigidity, thereby increasing the rate of vesicle rupture under the stress applied by DNA hybridization. In contrast, the presence of cholesterol had the opposite effect on the mechanical properties of the bilayer, and hence of the membrane fusion rates. The accessibility of receptor moieties (i.e., complementary DNA oligonucleotides) affected the fusion kinetics by modulating the rate of hybridization events. DNA accessibility was controlled by systematic variation of the length of the DNA receptor molecules and the thickness of the steric barrier comprised of adsorbed PEGylated lipids. These results provide design rules for understanding the trade-offs between response kinetics and other important system properties, such as nonspecific adsorption. Moreover, these findings improve our understanding of the biophysical properties of membrane fusion, an important process in both natural and model systems used for bioassay and bioimaging applications.