The delivery of genes or viruses using liposomes is a common approach used to enhance delivery efficiency. In the current study, to enhance delivery efficiency, proteoliposomes (PLs) containing adenovirus (Ad) were synthesized using dimyristoylphosphatidylcholine (DMPC), cholesterol, and apolipoprotein (apo) A-I. Wildtype-apoA-I (WT) or V156K-apoA-I (V156K) was then used as an apolipoprotein to compare the structural and functional differences of the PL. The particle diameter of V156K-PL-Ad was slightly larger than that of WT-PL-Ad based on native gel electrophoresis. V156K showed more rapid phospholipid bilayer formation than the WT based on DMPC-clearance. In addition, V156K exhibited a maximum fluorescence that was more blue than the WT in the PL state. Moreover, isothermal denaturation in response to the addition of guanidine hydrochloride (Gnd-HCl) revealed that V156K was more resistant, with no denaturation until 3 M Gnd-HCl was added. Additionally, electron microscopy revealed that the viral particle was well-associated with PL particles, which had a discoidal structure and a rouleax shape. In addition, treatment of Ad in the PL state showed enhanced GFP expression when compared totreatment with Ad alone or with DMPC-Ad in hepatoma and brain glioma cells. Cells treated with WT-PL-Ad and V156K-PL-Ad showed approximately 50% more GFP expression than cells treated with Ad alone or with DMPC-Ad after 24 hr of incubation at 37oC, indicating that viral stability was highly increased in the PL state. Furthermore, V156K-PL-Ad showed the highest expression of GFP in adult zebrafish (9 weeks old) at 5 days post-injection (10.5- and 3.8-fold more GFP expressed than Ad only and DMPC-Ad, respectively). In conclusion, the efficiency of viral delivery and the stability of the virus were significantly enhanced when the PL containing apoA-I was used in cellular and zebrafish models.