Carboxyl-terminal truncation of apolipoprotein B results in gradual loss of the ability to form buoyant lipoproteins in cultured human and rat liver cell lines
| Autor: | T C Jones, D.L. Graham, J Scott, Clive R. Pullinger, R J Pease, T J Knott |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Very low-density lipoprotein
Apolipoprotein B Gene Expression Lipoproteins VLDL Biology Transfection Biochemistry Infranatant Tumor Cells Cultured medicine Animals Humans Apolipoproteins B Gel electrophoresis DNA medicine.disease Precipitin Tests Molecular biology Rats Lipoproteins LDL Liver Mutation biology.protein lipids (amino acids peptides and proteins) Hypobetalipoproteinemia Chylomicron Lipoprotein |
| Zdroj: | Biochemistry. 30:5616-5621 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi00236a040 |
| Popis: | Apolipoprotein B has an obligatory role in the production of chylomicrons, VLDL, and LDL. Familial hypobetalipoproteinemia is a codominant disorder characterized by reduced levels of apo B containing lipoproteins in plasma. We have previously described mutations of the apo B gene in persons with hypobetalipoproteinemia that predict truncated forms of apo B designated apo B29 (1305 amino acid residues) and apo B39 (1799 residues). Apo B39 was present in the VLDL and LDL fractions of plasma, but apo B29 was not detected in the lipoprotein or infranatant fractions of plasma. Here we have investigated the regions of apo B necessary for apo B containing lipoprotein secretion by expression of constructs designed to express truncated forms of apo B. Apo B13 (583 residues), apo B17 (784 residues), apo B23 (1084 residues), apo B29 (1306 residues), and apo B41 (1880 residues) were transiently expressed in HepG2 cells, and apo B23 and apo B41 were stably expressed in McArdle 7777 cells. Lipoprotein (d less than 1.25 g/mL) and infranatant (d greater than 1.25 g/mL) fractions of conditioned medium were analyzed by immunoprecipitation and SDS-PAGE. The distribution between lipoprotein and infranatant fractions varied: apo B41 was found solely in the lipoprotein fraction; apo B29, apo B23, and apo B17 were present in both fractions, but with stepwise truncation, progressively more apo B was recovered in the infranatant; apo B13 was only in the infranatant. These results demonstrate that deletion from the carboxyl terminal of apo B41 results in a gradual loss of the ability of the truncated proteins to form buoyant lipoprotein particles. |
| Databáze: | OpenAIRE |
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