Characterization of the solubilized mosquito vitellogenin receptor

In this study we solubilized and characterized the receptor for the major egg yolk protein precursor, vitellogenin (Vg), from the yellow fever mosquito, Aedes aegypti. The receptor was solubilized from vitellogenic ovary membranes using octyl-β- -glucoside (OG). Under equilibrium binding conditions, ³⁵S]Vg bound with high affinity (K_d = 2.8 × 10^?8 M) to a single class of binding sites in solubilized ovary extracts. The solubilized receptor was present in ovarian extracts and bound selectively A. aegypti Vg and its storage form, vitellin (Vn). The receptor preparation was heat and trypsin sensitive. Binding of Vg to its receptor could be inhibited as well dissociated with suramin. The receptor was visualized by ligand-blotting as a 205 kDa protein under non-reducing conditions. It did not share immunological cross-reactivity with antibodies to chicken and locust Vg receptors. Vitellogenin, Vn and its purified subunits competed for binding to the receptor in the order, Vg ≈ Vn > Vn large subunit > Vn small subunit. Binding of dephosphorylated Vg was significantly reduced. Deglycosylated Vg, on the other hand, formed high molecular weight aggregates resulting in artifactually high binding which indicates importance of glycosylation for the stability of Vg molecule. During egg maturation, the number of receptor binding sites in ovaries correlated with the rate of Vg uptake and peaked between 24–30 h after which it reduced to no binding by 48 h post blood meal.