Murine mammary-derived cells secrete the N-terminal 41% of human apolipoprotein B on high density lipoprotein-sized lipoproteins containing a triacylglycerol-rich core.
The cDNA encoding the N-terminal 41% of human apolipoprotein B (apoB), apoB-41, was transfected into nonhepatic, nonintestinal, mammary-derived mouse cells (C127) to generate stably transfected cells expressing human apoB-41 (C127B-41). As determined by centrifugation, apoB-41 is secreted exclusively on lipoproteins (LPs) having a peak density of 1.13 g/ml. Electron microscopy of apoB-41-containing LPs purified by immunoaffinity chromatography showed round particles about 12 nm in diameter. No discoidal particles were observed. Characterization of apoB-41-associated lipids after labeling C127B-41 cells with [3H]oleate and immunoprecipitating the secreted LPs with antibodies to apoB showed that 3H-labeled triacylglycerols were a major lipid class and accounted for about 54% of the total labeled lipids. Cholesterol esters and phospholipids accounted for about 6% and 22%, respectively. Incubation of cells with 0.4 mM oleate resulted in an increased incorporation of the added oleate into lipids associated with secreted apoB-41, along with a 2- to 3-fold increased secretion of apoB-41. The newly formed LPs appear to be transported through the Golgi complex, as brefeldin A (1 microgram/ml) and monensin (1 microM) greatly reduced (> 90%) the secretion of labeled apoB-41 and the amount of triacylglycerol and phospholipid associated with it. Microsomal triacylglycerol transfer protein (MTP) was not detected in these cells. Taken together, the data presented demonstrate that apoB-41 can direct the assembly and secretion of LPs that contain a triacylglycerol-rich core in nonhepatic cells that apparently lack MTP. These cells, therefore, represent an important model for studying LP assembly and may offer some advantages over cultured hepatic or intestinal cells that express their endogenous apoB gene.
Herscovitz
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Department of Biophysics, Boston University School of Medicine, MA 02118-2394.
Kritis
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A
Talianidis
I
I
Zanni
E
E
Zannis
V
V
Small
D M
DM
D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D020126:Brefeldin A; D002352:Carrier Proteins / Q000032:analysis; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000032:analysis; D003517:Cyclopentanes / Q000494:pharmacology; D006023:Glycoproteins; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry / Q000378:metabolism*; D015674:Mammary Neoplasms, Animal; D051379:Mice; D008969:Molecular Sequence Data; D008985:Monensin / Q000494:pharmacology; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D010743:Phospholipids / Q000032:analysis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014280:Triglycerides / Q000032:analysis; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D003517:Cyclopentanes; D006023:Glycoproteins; D008075:Lipoproteins, HDL; D009829:Oleic Acids; D010743:Phospholipids; D011500:Protein Synthesis Inhibitors; D014280:Triglycerides; D020126:Brefeldin A; D019301:Oleic Acid; D008985:Monensin