Wax moth, Galleria mellonella fat body receptor for high-density lipophorin (HDLp)

摘要

To identify and characterize the HDLp (high-density lipophorin) receptor from Galleria mellonella (LpR(Gm)), we used techniques of ligand blotting. This method was, to our knowledge, first used to characterize the lipophorin receptor (LpR) in insects. LpR(Gm) had an approximate molecular weight of 97 kDa under non-reducing conditions and bound the HDLp specifically. The time-course of lipophorin binding to their receptor protein was rapid. The binding of lipophorins to their receptors was saturable with a K-d of 34.33 +/- 4.67 mug/ml. Although Ca2+ was essentially required in the binding of HDLp to their receptors, interestingly increasing concentration of Ca2+ has shown to have a slight inhibitory effect. EDTA was used here as Ca2+ chelating reagent, because Mg2+ in the binding buffer did not affect the binding of HDLp to their receptors, and inhibited the binding of HDLp and LpR(Gm) absolutely. Suramin (polysulfated polycyclic hydrocarbon), known to inhibit the binding of lipoproteins to their receptors, effectively abolished the binding of HDLp to their receptors. LpR(Gm) showed the stage specific binding activity especially in day 1-3 lost instar larval, prepupol, and day 1-3 adult stages.