Wax moth, Galleria mellonella fat body receptor for high-density lipophorin (HDLp).
To identify and characterize the HDLp (high-density lipophorin) receptor from Galleria mellonella (LpRGm), we used techniques of ligand blotting. This method was, to our knowledge, first used to characterize the lipophorin receptor (LpR) in insects. LpRGm 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 Kd of 34.33+/-4.67 microg/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 LpRGm absolutely. Suramin (polysulfated polycyclic hydrocarbon), known to inhibit the binding of lipoproteins to their receptors, effectively abolished the binding of HDLp to their receptors. LpRGm showed the stage specific binding activity especially in day 1-3 last instar larval, prepupal, and day 1-3 adult stages.
Lee
Chang Seok
CS
Laboratory of Animal Macromolecules, Physiology and Biochemistry, School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.
Han
Jik Hyon
JH
Lee
Sang Mong
SM
Hwang
Jae Sam
JS
Kang
Seok Woo
SW
Lee
Bong Hee
BH
Kim
Hak R
HR
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D002118:Calcium / Q000037:antagonists & inhibitors / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D005216:Fat Body / Q000378:metabolism*; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D009036:Moths / Q000378:metabolism*; D011869:Radioligand Assay; D018160:Receptors, Cytoplasmic and Nuclear / Q000302:isolation & purification / Q000378:metabolism*; D013498:Suramin / Q000494:pharmacology
D002352:Carrier Proteins; D002614:Chelating Agents; D019476:Insect Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D018160:Receptors, Cytoplasmic and Nuclear; C032830:lipophorin; C432945:lipophorin receptor; D013498:Suramin; D004492:Edetic Acid; D002118:Calcium