Microsomal triacylglycerol transfer protein prevents presecretory degradation of apolipoprotein B-100. A dithiothreitol-sensitive protease is involved.
The role of microsomal triacylglycerol transfer protein (MTP) in the secretion of apolipoprotein B-100 (apoB-100) has been studied using an inhibitor of MTP: 4'-bromo-3'-methylmetaqualone. In vitro, this compound inhibits trioleoylglycerol transfer between lipid vesicles mediated by MTP with an IC50 of 0.9 microM whereas it does not inhibit the lipid transfer mediated by the cholesteryl ester transfer protein. In HepG2 cells, 4'-bromo-3'-methylmetaqualone inhibits the secretion of apoB-100 with an IC50 of 0.3 microM, without affecting the secretion of several other proteins like apoA-I or albumin. Moreover, there is no accumulation of apoB-100 in treated cells. Oleic acid, which increases apoB-100 secretion, only slightly modifies the IC50 of 4'-bromo-3'-methylmetaqualone (0.5 microM). The latter has no effect on the synthesis of major lipids within the cell, but decreases the secretion of triacylglycerol into apoB-100-containing lipoproteins. Pulse/chase experiments reveal that 4'-bromo-3'-methylmetaqualone acts on apoB-100 production either at the co-translational or post-translational level. The cysteine protease inhibitor N-acetyl-leucyl-leucyl-norleucinal does not protect apoB-100 from the 4'-bromo-3'-methylmetaqualone effect but seems to be involved in a later step of apoB-100 intracellular degradation. By contrast, dithiothreitol can totally reverse the effect of the MTP inhibitor on apoB-100 production. The mechanism of MTP-mediated lipid assembly with apoB-100 is discussed.
Benoist
F
F
Laboratorie Glaxo Wellcome, Centre de Recherche, France.
Nicodeme
E
E
Grand-Perret
T
T
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000378:metabolism*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002460:Cell Line; D053480:Cholesterol Ester Transfer Proteins; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D006023:Glycoproteins; D006801:Humans; D066298:In Vitro Techniques; D007976:Leupeptins / Q000494:pharmacology; D050356:Lipid Metabolism; D008055:Lipids / Q000096:biosynthesis; D008702:Methaqualone / Q000031:analogs & derivatives / Q000494:pharmacology; D008861:Microsomes / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism*
C101933:4'-bromo-3'-methylmethaqualone; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins; D007976:Leupeptins; D008055:Lipids; D014280:Triglycerides; C063261:acetylleucyl-leucyl-norleucinal; D008702:Methaqualone; D010450:Endopeptidases; D004229:Dithiothreitol