Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Apr;87(7):2390–2394. doi: 10.1073/pnas.87.7.2390

Possible role of the Golgi apparatus in the assembly of very low density lipoprotein.

M J Bamberger 1, M D Lane 1
PMCID: PMC53694  PMID: 2181435

Abstract

Transit of newly synthesized triacyl[3H]-glycerol through organelles of the secretory system leading to assembly into nascent very low density lipoproteins (VLDLs) or to cytoplasmic storage was investigated in chick hepatocytes. Cells in monolayer culture were pulse-labeled with [2-3H]glycerol, and after different periods of chase with unlabeled glycerol, the movement of triacyl[3H]glycerol through the endoplasmic reticulum (ER) and Golgi and the incorporation into nascent VLDL and cytoplasmic triacylglycerol-rich vesicles (TGRVs) were determined. Initially, newly synthesized triacyl[3H]glycerol is tightly associated with the ER. Movement from the ER of triacyl[3H]glycerol destined for cytoplasmic storage (as TGRVs) is extremely rapid and virtually complete within 8 min of chase. After 8 min of chase, triacyl[3H]glycerol lost from organelles of the secretory system was accounted for entirely as triacyl[3H]glycerol secreted as VLDL. Comparison of the rates of movement of triacyl[3H]glycerol, apolipoprotein B, apolipoprotein II, and apolipoprotein A-I through the ER and Golgi and of their secretion in nascent VLDL suggests that assembly of triacyglycerol with apolipoproteins occurs in the Golgi. Experiments with permeabilized hepatocytes supplemented with cytosol show that newly synthesized triacyl[3H]glycerol and [3H]phospholipid moves from the ER through the full-density range of Golgi elements and is dependent upon supplementary ATP.

Full text

PDF
2390

Images in this article

2392Image
on p.2392
2393Image
on p.2393
2393Image
on p.2393

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  2. Balch W. E., Rothman J. E. Characterization of protein transport between successive compartments of the Golgi apparatus: asymmetric properties of donor and acceptor activities in a cell-free system. Arch Biochem Biophys. 1985 Jul;240(1):413–425. doi: 10.1016/0003-9861(85)90046-3. [DOI] [PubMed] [Google Scholar]
  3. Bamberger M. J., Lane M. D. Assembly of very low density lipoprotein in the hepatocyte. Differential transport of apoproteins through the secretory pathway. J Biol Chem. 1988 Aug 25;263(24):11868–11878. [PubMed] [Google Scholar]
  4. Beckers C. J., Keller D. S., Balch W. E. Semi-intact cells permeable to macromolecules: use in reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex. Cell. 1987 Aug 14;50(4):523–534. doi: 10.1016/0092-8674(87)90025-0. [DOI] [PubMed] [Google Scholar]
  5. Bell R. M., Coleman R. A. Enzymes of glycerolipid synthesis in eukaryotes. Annu Rev Biochem. 1980;49:459–487. doi: 10.1146/annurev.bi.49.070180.002331. [DOI] [PubMed] [Google Scholar]
  6. Boström K., Wettesten M., Borén J., Bondjers G., Wiklund O., Olofsson S. O. Pulse-chase studies of the synthesis and intracellular transport of apolipoprotein B-100 in Hep G2 cells. J Biol Chem. 1986 Oct 15;261(29):13800–13806. [PubMed] [Google Scholar]
  7. Hoeg J. M., Meng M. S., Ronan R., Fairwell T., Brewer H. B., Jr Human apolipoprotein A-I. Post-translational modification by fatty acid acylation. J Biol Chem. 1986 Mar 25;261(9):3911–3914. [PubMed] [Google Scholar]
  8. Janero D. R., Lane M. D. Sequential assembly of very low density lipoprotein apolipoproteins, triacylglycerol, and phosphoglycerides by the intact liver cell. J Biol Chem. 1983 Dec 10;258(23):14496–14504. [PubMed] [Google Scholar]
  9. Kane J. P. Apolipoprotein B: structural and metabolic heterogeneity. Annu Rev Physiol. 1983;45:637–650. doi: 10.1146/annurev.ph.45.030183.003225. [DOI] [PubMed] [Google Scholar]
  10. Law S. W., Grant S. M., Higuchi K., Hospattankar A., Lackner K., Lee N., Brewer H. B., Jr Human liver apolipoprotein B-100 cDNA: complete nucleic acid and derived amino acid sequence. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8142–8146. doi: 10.1073/pnas.83.21.8142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mooney R. A., Lane M. D. Control of ketogenesis and fatty-acid synthesis at the mitochondrial branch-point for acetyl-CoA in the chick liver cell: effect of adenosine 3',5'-monophosphate. Eur J Biochem. 1982 Jan;121(2):281–287. doi: 10.1111/j.1432-1033.1982.tb05783.x. [DOI] [PubMed] [Google Scholar]
  12. Mooney R. A., Lane M. D. Formation and turnover of triglyceride-rich vesicles in the chick liver cell. Effects of cAMP and carnitine on triglyceride mobilization and conversion to ketones. J Biol Chem. 1981 Nov 25;256(22):11724–11733. [PubMed] [Google Scholar]
  13. Siuta-Mangano P., Howard S. C., Lennarz W. J., Lane M. D. Synthesis, processing, and secretion of apolipoprotein B by the chick liver cell. J Biol Chem. 1982 Apr 25;257(8):4292–4300. [PubMed] [Google Scholar]
  14. Siuta-Mangano P., Janero D. R., Lane M. D. Association and assembly of triglyceride and phospholipid with glycosylated and unglycosylated apoproteins of very low density lipoprotein in the intact liver cell. J Biol Chem. 1982 Oct 10;257(19):11463–11467. [PubMed] [Google Scholar]
  15. Siuta-Mangano P., Lane M. D. Very low density lipoprotein synthesis and secretion. Extrusion of apoprotein B nascent chains through the membrane of the endoplasmic reticulum without protein synthesis. J Biol Chem. 1981 Mar 10;256(5):2094–2097. [PubMed] [Google Scholar]
  16. Tarlow D. M., Watkins P. A., Reed R. E., Miller R. S., Zwergel E. E., Lane M. D. Lipogenesis and the synthesis and secretion of very low density lipoprotein by avian liver cells in nonproliferating monolayer culture. Hormonal effects. J Cell Biol. 1977 May;73(2):332–353. doi: 10.1083/jcb.73.2.332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. ungenberg de Jong J. J., Marsh J. B. Biosynthesis of plasma lipoproteins by rat liver ribosomes. J Biol Chem. 1968 Jan 10;243(1):192–199. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES