Effects of temperature and lipid composition on the serum albumin-induced aggregation and fusion of small unilamellar vesicles.
Small unilamellar vesicles of egg phosphatidylcholine (PC) or dimyristoylphosphatidylcholine, mixed with small unilamellar vesicles labelled with 2-(10-(1-pyrene)decanoyl)phosphatidylcholine, exhibit a constant average size and excimer to monomer (E/M) ratio for several hours when incubated at pH 3.6 at a temperature higher than the phase transition temperature (Tc) of the lipids. Addition of bovine serum albumin to this system produces a transient turbidity increase, a fast decrease in the E/M ratio, a partial loss of vesicle-entrapped [14C]sucrose and a measurable leak-in of externally added sucrose. Sepharose 4B filtration of the system demonstrates that the E/M ratio decrease is strictly paralleled by the formation of liposomes which exhibit a low E/M ratio and a hydrodynamic radius larger than that of small unilamellar vesicles. These data demonstrate that the E/M ratio decrease can be unequivocally ascribed to a vesicle-vesicle fusion process induced by serum albumin. The rate of serum-albumin induced fusion of small unilamellar vesicles is: (a) maximal at a stoichiometric ratio of approx. 2 albumins per vesicle; (b) sensitive to the nature of the lipid and; (c) not altered when human serum albumin replaces bovine serum albumin. The rate of albumin-induced fusion of dimyristoylphosphatidylcholine small unilamellar vesicles is higher below the Tc of the lipid and increases with temperature above the Tc. The formation of protein-bound aggregates with defined stoichiometries and a high local vesicle concentration, as well as changes in the local degree of hydration, are proposed to be the driving forces for the protein-induced vesicle fusion in this system.
D000818:Animals; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine; D005456:Fluorescent Dyes; D007700:Kinetics; D008081:Liposomes; D008246:Lysophospholipids; D010713:Phosphatidylcholines; D012710:Serum Albumin, Bovine; D013053:Spectrophotometry; D013395:Sucrose; D013816:Thermodynamics
C033518:2-(10-(1-pyrene)decanoyl)phosphatidylcholine; D005456:Fluorescent Dyes; D008081:Liposomes; D008246:Lysophospholipids; D010713:Phosphatidylcholines; D012710:Serum Albumin, Bovine; D013395:Sucrose; D004134:Dimyristoylphosphatidylcholine