Identification of a functional receptor differing from the LDL receptor that catabolizes chylomicron remnant in Hep G2 cells.
We investigated types of lipoprotein receptors on Hep G2 cells using a monoclonal antibody against the LDL receptor. IgG-C7 inhibited the binding and internalization of 125I-labeled low density lipoprotein (LDL) in Hep G2 cells with upregulated and downregulated LDL receptors by 90% of control values. Binding and internalization of 125I-labeled chylomicron remnant in Hep G2 cells with upregulated and downregulated LDL receptors was 50% and 85%, respectively, of control values after exposure to IgG-C7. Excess unlabeled chylomicron remnant inhibited binding and internalization of 125I-labeled chylomicron remnant in Hep G2 cells with downregulated LDL receptors completely. Pronase treatment abolished binding and internalization of 125I-labeled LDL and 125I-labeled chylomicron remnant in Hep G2 cells. When solubilized fractions of Hep G2 cells were immunoprecipitated with IgG-C7, the binding activity of 125I-labeled chylomicron remnant to reconstituted vesicles was unchanged. 45Ca blotting analysis showed the presence of 45Ca binding protein (approximately 600 kDa) in Hep G2 cells. The amount of 45Ca binding protein was not affected by cholesterol and was abolished by pronase treatment. These results suggest the existence of a functional receptor other than the LDL receptor that catabolizes chylomicron remnant in Hep G2 cells and that this receptor may correspond to LDL receptor-related protein.
Hayashi
K
K
First Department of Internal Medicine, Hiroshima University School of Medicine, Japan.
Nakashima
K
K
Saeki
M
M
Kurushima
H
H
Kurokawa
J
J
Kuga
Y
Y
Nomura
S
S
Ohkura
Y
Y
Ohtani
H
H
Tanaka
K
K
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002914:Chylomicrons / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008071:Lipoprotein Lipase / Q000494:pharmacology; D011402:Pronase / Q000494:pharmacology; D011869:Radioligand Assay; D011973:Receptors, LDL / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002135:Calcium-Binding Proteins; D002914:Chylomicrons; D011973:Receptors, LDL; D008071:Lipoprotein Lipase; D011402:Pronase