Decreased secretion of ApoB follows inhibition of ApoB-MTP binding by a novel antagonist.
Apolipoprotein B (apoB) and microsomal triglyceride transfer protein (MTP) are essential for the efficient assembly of triglyceride-rich lipoproteins. Evidence has been presented for physical interactions between these proteins. To study the importance of apoB-MTP binding in apoB secretion, we have identified a compound, AGI-S17, that inhibited (60-70% at 40 microM) the binding of various apoB peptides to MTP but not to an anti-apoB monoclonal antibody, 1D1, whose epitope overlaps with an MTP binding site in apoB. AGI-S17 had no significant effect on the lipid transfer activity of the purified MTP. In contrast, another antagonist, BMS-200150, did not affect apoB-MTP binding but inhibited MTP's lipid transfer activity. The differential effects of these inhibitors suggest two functionally independent, apoB binding and lipid transfer, domains in MTP. AGI-S17 was then used to study its effect on the lipid transfer and apoB binding activities of MTP in HepG2 cells. AGI-S17 had no effect on cellular lipid transfer activities, but it inhibited coimmunoprecipitation of apoB with MTP. These studies indicate that AGI-S17 inhibits apoB-MTP binding but has no effect on MTP's lipid transfer activity. Experiments were then performed to study the effect of inhibition of apoB-MTP binding on apoB secretion in HepG2 cells. AGI-S17 (40 microM) did not affect cell protein levels but decreased the total mass of apoB secreted by 70-85%. Similarly, AGI-S17 inhibited the secretion of nascent apoB by 60-80%, but did not affect albumin secretion. These studies indicate that AGI-S17 decreases apoB secretion most likely by inhibiting apoB-MTP interactions. Thus, the binding of MTP to apoB may be important for the assembly and secretion of apoB-containing lipoproteins and can be a potential target for the development of lipid-lowering drugs. It is proposed that the apoB binding may represent MTP's chaperone activity that assists in the transfer from the membrane to the lumen of the endoplasmic reticulum and in the net lipidation of nascent apoB, and may be essential for lipoprotein assembly and secretion.
Bakillah
A
A
Department of Pathology, School of Medicine, MCP Hahnemann University, Philadelphia, Pennsylvania 19129, USA.
Nayak
N
N
Saxena
U
U
Medford
R M
RM
Hussain
M M
MM
D000418:Albumins / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology; D001055:Apolipoproteins B / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism*; D001665:Binding Sites / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D054833:Isoindoles; D050356:Lipid Metabolism; D008954:Models, Biological; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D010880:Piperidines / Q000494:pharmacology; D011233:Precipitin Tests; D011485:Protein Binding / Q000187:drug effects; D014407:Tumor Cells, Cultured
D000418:Albumins; D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; C101970:BMS 200150; D002352:Carrier Proteins; D007211:Indoles; D054833:Isoindoles; D010446:Peptide Fragments; D010880:Piperidines; C106092:microsomal triglyceride transfer protein