Immunoenzyme assessment of human apoB-lipoprotein binding to immobilized receptor of low density lipoproteins. 2. Binding of isolated lipoproteins.
The receptor of low density lipoproteins (LDL-receptor) from bovine adrenal cortex membranes was immobilized in standard 96-well polystyrene plates using monoclonal V5-antibodies to the LDL-receptor. The binding of the immobilized LDL-receptor with human low density lipoproteins (LDL) and very low density lipoproteins (VLDL) was determined using peroxidase-labelled antibodies to human apoB. The value of Kd for the interaction of LDL with the immobilized LDL-receptor for 40 samples of LDL was found to be from 5 to 20 micrograms apoB per ml. The immobilized LDL-receptor failed to bind LDL modified by acetylation or malonic dialdehyde, while the binding of non-modified LDL to the immobilized LDL-receptor was inhibited in the presence of EDTA, which is known to be specific for the interaction of LDL with the LDL-receptor. Unlike LDL, VLDL were more variable in the binding to the LDL-receptor. The value of Kd for the interaction of VLDL with the LDL-receptor for 40 samples of VLDL was found to be from 0.5 to 10 micrograms apoB per ml. Thus, the described method is suggested to study the interaction of apoB-containing lipoproteins with the LDL-receptor.
Tsibulsky
V P
VP
Institute of Experimental Cardiology, Russian Cardiology Complex, Moscow, Russia.
Yakushkin
V V
VV
Preobrazhensky
S N
SN
D000107:Acetylation; D000302:Adrenal Cortex / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B / Q000378:metabolism*; D002417:Cattle; D004492:Edetic Acid; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D007700:Kinetics; D008074:Lipoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008079:Lipoproteins, VLDL / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008315:Malondialdehyde; D051379:Mice; D011485:Protein Binding; D011973:Receptors, LDL / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D008074:Lipoproteins; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D011973:Receptors, LDL; D008315:Malondialdehyde; D004492:Edetic Acid