The hydatid disease parasite Echinococcus granulosus has a restricted lipid metabolism, and needs to harvest essential lipids from the host. Antigen B (EgAgB), an abundant lipoprotein of the larval stage (hydatid cyst), is thought to be important in lipid storage and transport. It contains a wide variety of lipid classes, from highly hydrophobic compounds to phospholipids. Its protein component belongs to the cestode-specific Hydrophobic Ligand Binding Protein family, which includes five 8-kDa isoforms encoded by a multigene family (EgAgB1-EgAgB5). How lipid and protein components are assembled into EgAgB particles remains unknown. EgAgB apolipoproteins self-associate into large oligomers, but the functional contribution of lipids to oligomerization is uncertain. Furthermore, binding of fatty acids to some EgAgB subunits has been reported, but their ability to bind other lipids and transfer them to acceptor membranes has not been studied.
Lipid-free EgAgB subunits obtained by reverse-phase HPLC were used to analyse their oligomerization, ligand binding and membrane interaction properties. Size exclusion chromatography and cross-linking experiments showed that EgAgB8/2 and EgAgB8/3 can self-associate, suggesting that lipids are not required for oligomerization. Furthermore, using fluorescent probes, both subunits were found to bind fatty acids, but not cholesterol analogues. Analysis of fatty acid transfer to phospholipid vesicles demonstrated that EgAgB8/2 and EgAgB8/3 are potentially capable of transferring fatty acids to membranes, and that the efficiency of transfer is dependent on the surface charge of the vesicles.
We show that EgAgB apolipoproteins can oligomerize in the absence of lipids, and can bind and transfer fatty acids to phospholipid membranes. Since imported fatty acids are essential for Echinococcus granulosus, these findings provide a mechanism whereby EgAgB could engage in lipid acquisition and/or transport between parasite tissues. These results may therefore indicate vulnerabilities open to targeting by new types of drugs for hydatidosis therapy.
Echinococcus granulosus is a causative agent of hydatidosis, a parasitic disease that affects humans and livestock with significant economic and public health impact worldwide. Antigen B (EgAgB), an abundant product of E. granulosus larvae, is a lipoprotein that carries a wide variety of lipids, including fatty acids and cholesterol. As E. granulosus is unable to synthesize these lipids, EgAgB likely plays an important role in parasite metabolism, participating in both the acquisition of host lipids and their distribution between parasite tissues. The protein component of EgAgB consists of 8 kDa subunits encoded by separate genes. However, the biochemical properties of EgAgB subunits, particularly their ability to bind and transfer lipids, are poorly known. Herein, using in vitro assays, we found that EgAgB subunits were capable of oligomerizing in the absence of lipids and to bind fatty acids, but not cholesterol. Moreover, EgAgB subunits showed the ability to transfer fatty acids to artificial phospholipid membranes. These results indicate new points of attack at which the parasite might be vulnerable to drugs.