Specific binding of avidin to biotin containing lipid lamella surfaces studied with monolayers and liposomes

The interaction of avidin (from egg white) with phospholipid (monolayer and bilayer) model membranes containing biotin-conjugated phospholipids has been studied. In the first part, using surface sensitive techniques (ellipsometry and surface plasmon resonance) we demonstrated that the nonspecific adsorption of avidin to phospholipid lamella could be abolished by adding an amount of Ca²⁺, Mg²⁺, or Ba²⁺ that led to an electrostatic interaction. The specific binding of avidin to lipid mixtures containing biotin-conjugated phospholipids was obviously composition dependent. The ratio 1:12 of a B-DPPE/DPPE mixture was found to be the optimum molar ratio. When we compared the results from the surface sensitive techniques with those from the electron micrographs of a two dimensional crystal of avidin (obtained in our laboratory), the optimum ratio was found to be determined by the effect of lateral steric hindrance. In the second part, we observed the pattern of the layers of fluorescently labeled phospholipid and adsorbed proteins with a home-made micro fluorescence film balance. The fluorescence images showed that avidin was preferentially bound to the receptors that were in the fluid domains. Further, with a sensitive fluoresence assay method, the effect of the phase behavior of liposomes on the specific binding of avidin was measured. This showed that avidin interacted with biotin-lipid more weakly in the gel state liposome than in the liquid state liposome. The major conclusion was that the binding of avidin to a membrane bound model receptor was significantly restricted by two factors: one was the lateral steric hindrance and the other was the fluidity of the model membrane.Abbreviations B-DPPE Biotinyl dipalmitoylphosphatidyl ethanolamine - B-DMPE Biotinyl dimyristoylphosphatidyl ethanolamine - BNHS d-biotin-N-hydroxysuccinimide ester - DMPA dimyristoylphosphatidyl acid - DMPC dimyristoylphosphatidyl choline - DMPS dimyristoylphosphatidyl serine - DOPC dioleoylphosphatidyl choline - DPPC dipalmitoylphosphatidyl choline - DPPE dipalmitoylphosphatidyl ethanolamine - FITC fluorescein isothiocyanate - RDB-DOPE N(Lissamine rhodamine B sulfonyl) dioleoyl phosphatidylethanolamine - SPR surface plasmon resonanceCorrespondence to: S. F. Sui