Previous studies show that translocation and degradation of apolipoprotein B (apoB), two processes occurring on or within the endoplasmic reticulum, determine how much de novo synthesized apoB is secreted. We determined which of these processes regulates the intracellular fate of apoB by examining whether degradation determines how much apoB is translocated or if translocation determines how much apoB is degraded. HepG2 cells, treated with the cysteine active site protease inhibitor ALLN, previously shown to block the degradation of translocation-arrested apoB in Chinese hamster ovary cells (Du, E., Kurth, J., Wang, S.-L., Humiston, P., and Davis, R. A.(1994) J. Biol. Chem. 269, 24169-24176), showed a 10-fold increase in the accumulation of de novo synthesized [S]methionine-labeled apoB. The majority (80%) of the apoB accumulated in response to ALLN was in the microsomal fraction. In contrast, ALLN did not effect apoB secretion. Since ALLN did not effect the intracellular accumulation of [S]methionine-labeled albumin and other proteins (trichloroacetic acid-precipitable [S]methionine-labeled proteins), its effect on apoB was specific. Pulse-chase studies showed that ALLN dramatically reduced the first-order rate of removal of [S]methionine-labeled apoB from the cell but did not effect its rate of secretion. The finding that ALLN caused the intracellular accumulation of incompletely translated chains of apoB suggests that at least some of the degradation occurs at the ribosomal level. Moreover, 85% of the apoB that accumulated in isolated microsomes in response to ALLN was accessible to exogenous trypsin, indicating this pool of apoB was incompletely translocated. The combined data suggest that translocation, not degradation, determines the intracellular fate of de novo synthesized apoB.

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