Degradation of newly synthesized apolipoprotein B-100 in a pre-Golgi compartment.
The synthesis and secretion of apolipoprotein (apo) B-100 have been studied in a human hepatoblastoma cell line, the Hep G2 cells. Pulse-chase analysis showed that apoB-100 was not quantitatively recovered in the culture medium. To reveal the intracellular degradation of apoB-100 prior to secretion, cells were incubated with 1 microgram/ml Brefeldin A (BFA) which impeded protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus and the fate of apoB-100 retained in the cells was traced at 37 degrees C. A significant amount of intracellular apoB-100 (40-60%/h) was degraded during the chase period, whereas apoA-1 remained intact. ApoB-100 degradation was temperature dependent, no degradation was observed below 20 degrees C. This degradation process was not inhibited by chloroquine, leupeptin, pepstatin, and chymostatin, suggesting that lysosomal proteases were not involved and that apoB-100 was degraded in a pre-Golgi compartment which is either part of, or closely related to, the ER. Preincubation of cells with low density lipoproteins (LDL) induced a 22-32% increase in the degradation of apoB-100. This result raised the possibility that secretion of apoB-100 might be regulated through the intracellular degradation of apoB-100. These results suggest the existence of the degradation pathway for apoB-100 in a pre-Golgi compartment and an unique regulatory mechanism for apoB-100 secretion.
Sato
R
R
Department of Microbiology and Molecular Pathology, Teikyo University, Kanagawa, Japan.
Imanaka
T
T
Takatsuki
A
A
Takano
T
T
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D016632:Apolipoprotein A-I; D053299:Apolipoprotein B-100; D001054:Apolipoproteins A / Q000378:metabolism; D001055:Apolipoproteins B / Q000096:biosynthesis / Q000378:metabolism*; D020126:Brefeldin A; D002460:Cell Line; D003517:Cyclopentanes / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D013462:Sulfur Radioisotopes; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis
D000900:Anti-Bacterial Agents; D016632:Apolipoprotein A-I; D053299:Apolipoprotein B-100; D001054:Apolipoproteins A; D001055:Apolipoproteins B; D003517:Cyclopentanes; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D013462:Sulfur Radioisotopes; D000515:alpha 1-Antitrypsin; D020126:Brefeldin A; D008715:Methionine