Binding of locust high-density lipophorin to fat body proteins monitored by an enzyme-linked immunosorbant assay.
Binding of locust high-density lipophorin (HDLp) to fat body proteins coated on immunoassay plates was studied using the ELISA method and ligand blotting techniques. HDLp binding proved to be correlated with the amount of fat body protein coated. From the concentration-dependent total HDLp binding an equilibrium dissociation constant could be calculated (Kd = 1.6 x 10(-8) M). Heparin inhibits the HDLp binding, indicating that positively charged groups are involved in the HDLp-fat body interaction. These groups were shown to be arginyl residues, as the arginine-specific treatment of HDLp by 1,2-cyclohexanedione resulted in a approximately 50% decrease in the binding ability of HDLp. HDLp binding is also affected by the pH. A decrease from pH 7.5 to pH 6.5 increases the binding affinity by approximately 250%. A monoclonal antibody specific for apolipophorin-II (apoLp-II) hampered the HDLp binding significantly, whereas a monoclonal anti-apoLp-I had no effect. Locust fat body HDLp binding proteins are highly specific for locust HDLp.
Schulz
T K
TK
Department of Experimental Zoology, University of Utrecht.
Van der Horst
D J
DJ
Beenakkers
A M
AM
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001059:Apoproteins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D003512:Cyclohexanones / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005216:Fat Body / Q000378:metabolism*; D006110:Grasshoppers / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008074:Lipoproteins; D011485:Protein Binding / Q000187:drug effects; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits
D000911:Antibodies, Monoclonal; D001059:Apoproteins; D002352:Carrier Proteins; D003512:Cyclohexanones; D008074:Lipoproteins; D011506:Proteins; C032830:lipophorin; C018031:1,2-cyclohexanedione; D006493:Heparin