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The assembly of hepatic very low density lipoproteins: Evidence of a role for the Golgi apparatus

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Lipids

Abstract

Previous studies from our laboratory have suggested that the assembly of lipoproteins by the liver is not completed in the rough endoplasmic reticulum but continues while the particles are en route to or within the Golgi apparatus. To investigate further the role of the Golgi apparatus in lipoprotein assembly, mice were injected with [3H]glycerol and killed 7.5 to 45 min after injection. Microsomes and Golgi apparatus-rich fractions were isolated from the livers and separated into membrane and content fractions. TG within microsomal and Golgi membranes were labeled, rapidly reaching peak specific activity within 7.5 min of isotope injection. The specific activity of TG in microsomal membranes decreased to approximately 40% of peak values by 45 min, whereas the specific activity of TG in the Golgi membranes decreased to approximately 30% of peak values by 45 min. To determine whether the turnover of the Golgi membrane TG pool was dependent on microsomal TG transfer protein (MTP), mice were gavaged with an MTP inhibitor, and the labeling experiments were repeated. Inhibition of MTP attenuated the turnover of newly synthesized Golgi membrane TG by approximately 50% and the turnover of microsomal membrane TG by approximately 40%. Based on the rapid turnover of the Golgi membrane TG pool and the attenuation of the turnover of this pool by MTP inhibitor, we propose that lipid is transferred from the Golgi membrane to luminal lipoproteins in an MTP-dependent manner. The results support our hypothesis that assembly of VLDL continues within the Golgi apparatus.

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Abbreviations

apoB:

apolipoprotein B

DGAT:

DAG acyltransferase

ER:

endoplasmic reticulum

MTP:

microsomal TG transfer protein

References

  1. Alexander, C.A., Hamilton, R.L., and Havel, R.J. (1976) Subcellular Localization of B Apoprotein of Plasma Lipoproteins in Rat Liver, J. Cell Biol. 69, 241–263.

    Article  PubMed  CAS  Google Scholar 

  2. Boren, J.S., Rustaeus, S., and Olofsson, S.-O. (1994) Studies on the Assembly of Apolipoprotein B-100 and B-48-Containing Very Low Density Lipoproteins in McA-RH7777 Cells, J. Biol. Chem. 269, 25879–25888.

    PubMed  CAS  Google Scholar 

  3. Olofsson, S.-O., Asp, L., and Borén, J. (1999) The Assembly and Secretion of Apolipoprotein B-Containing Lipoproteins, Curr. Opin. Lipidol. 10, 341–346.

    Article  PubMed  CAS  Google Scholar 

  4. Bamberger, M., and Lane, M. (1990) Possible Role of the Golgi Apparatus in the Assembly of Very Low Density Lipoprotein, Proc. Natl. Acad. Sci. USA 87, 2390–2394.

    Article  PubMed  CAS  Google Scholar 

  5. Higgins, J. (1998) Evidence That During Very Low Density Lipoprotein Assembly in Rat Hepatocytes Most of the Triacylglycerol and Phospholipid Are Packaged with Apolipoprotein B in the Golgi Complex, FEBS Lett. 232, 405–408.

    Article  Google Scholar 

  6. Boström, K., Borén, J., Wettesten, M., Sjöberg, A., Bondjers, G., Wiklund, O., Carlsson, P., and Olofsson, S.-O. (1988) Studies on the Assembly of ApoB-100-Containing Lipoproteins in HepG2 Cells, J. Biol. Chem. 263, 4434–4442.

    PubMed  Google Scholar 

  7. Cartwright, I., and Higgins, J. (1995) Intracellular Events in the Assembly of Very Low Density Lipoprotein Lipids with Apolipoprotein B in Isolated Rabbit Hepatocytes, Biochem. J. 310, 897–907.

    PubMed  CAS  Google Scholar 

  8. Stillemark, P., Borén, J., Andersson, M., Larsson, T., Rustaeus, S., Karlsson, K.-A., and Olofsson, S.-O. (2000) The Assembly and Secretion of Apolipoprotein B-48-Containing Very Low Density Lipoproteins in McA-RH7777 Cells, J. Biol. Chem. 275, 10506–10513.

    Article  PubMed  CAS  Google Scholar 

  9. Swift, L.L. (1995) The Assembly of Very Low Density Lipoprotein in Rat Liver: A Study of Nascent Particles Recovered from the Rough Endoplasmic Reticulum, J. Lipid Res. 36, 395–406.

    PubMed  CAS  Google Scholar 

  10. Swift, L.L., Valyi-Nagy, K., Rowland, C., and Harris, C. (2001) Assembly of Very Low Density Lipoproteins in Mouse Liver: Evidence for Heterogeneity of Particle Density in the Golgi Apparatus, J. Lipid Res. 42, 218–224.

    PubMed  CAS  Google Scholar 

  11. Fujiki, Y., Hubbard, A.L., Fowler, S., and Lazarow, P.B. (1982) Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum, J. Cell Biol. 93, 97–102.

    Article  PubMed  CAS  Google Scholar 

  12. Swift, L.L., Farkas, M.H., Major, A.S., Valyi-Nagy, K., Linton, M.F., and Fazio, S. (2001) A Recycling Pathway for Resecretion of Internalized Apolipoprotein E in Liver Cells, J. Biol. Chem. 276, 22965–22970.

    Article  PubMed  CAS  Google Scholar 

  13. Folch, J., Lees, M., and Sloane Stanley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues, J. Biol. Chem. 226, 497–509.

    PubMed  CAS  Google Scholar 

  14. Bersot, T., Brown, W., Levy, R., Windmueller, H., Fredrickson, D., and LeQuire, V. (1970) Further Characterization of the Apolipoproteins of Rat Plasma Lipoproteins, Biochemistry 9, 3427–3433.

    Article  PubMed  CAS  Google Scholar 

  15. Dittmer, J., and Wells, M. (1969) Quantitative and Qualitative Analysis of Lipids and Lipid Components, Methods Enzymol. 14, 482–530.

    Article  CAS  Google Scholar 

  16. Morton, R., and Evans, T. (1992) Modification of the Bicinchoninic Acid Protein Assay to Eliminate Lipid Interference in Determining Lipoprotein Protein Content, Anal. Biochem. 204, 332–334.

    Article  PubMed  CAS  Google Scholar 

  17. Asp, L., Claesson, C., Borén, J., and Olofsson, S.-O. (2000) ADP-Ribosylation Factor 1 and Its Activation of Phospholipase D Are Important for the Assembly of Very Low Density Lipoproteins, J. Biol. Chem. 275, 26285–26292.

    Article  PubMed  CAS  Google Scholar 

  18. Hebbachi, A.-M., and Gibbons, G. (1999) Inactivation of Microsomal Triglyceride Transfer Protein Impairs the Normal Redistribution but Not the Turnover of Newly Synthesized Glycerolipid in the Cytosol, Endoplasmic Reticulum and Golgi of Primary Rat Hepatocytes, Biochim. Biophys. Acta 1441, 36–50.

    PubMed  CAS  Google Scholar 

  19. Gibbons, G.F., and Wiggins, D. (1995) Intracellular Triacylglycerol Lipase: Its Role in the Assembly of Hepatic Very-Low-Density Lipoprotein (VLDL), Adv. Enzyme Regul. 35, 179–198.

    Article  PubMed  CAS  Google Scholar 

  20. Salter, A.M., Wiggins, D., Sessions, V.A., and Gibbons, G.F. (1998) The Intracellular Triacylglycerol/Fatty Acid Cycle: A Comparison of Its Activity in Hepatocytes Which Secrete Exclusively Apolipoprotein (apo) B100 Very-Low-Density Lipoprotein (VLDL) and in Those Which Secrete Predominantly apoB48 VLDL, Biochem. J. 332, 667–672.

    PubMed  CAS  Google Scholar 

  21. Gibbons, G.F., Islam, K., and Pease, R.J. (2000) Mobilisation of Triacylglycerol Stores, Biochim. Biophys. Acta 1483, 37–57.

    PubMed  CAS  Google Scholar 

  22. Bannykh, S., and Balch, W. (1998) Selective Transport of Cargo Between the Endoplasmic Reticulum and Golgi Compartments, Histochem. Cell Biol. 109, 463–475.

    Article  PubMed  CAS  Google Scholar 

  23. Palade, G.E. (1975) Intracellular Aspects of the Process of Protein Transport, Science 189, 347–354.

    Article  PubMed  CAS  Google Scholar 

  24. Moreau, P., Rodriguez, M., Cassagne, C., Morré, D.M., and Morré, D.J. (1991) Trafficking of Lipids from the Endoplasmic Reticulum to the Golgi Apparatus in a Cell-Free System from Rat Liver, J. Biol. Chem. 266, 4322–4328.

    PubMed  CAS  Google Scholar 

  25. Hebbachi, A.-M., Brown, A.-M., and Gibbons, G. (1999) Suppression of Cytosolic Triacylglycerol Recruitment for Very Low Density Lipoprotein Assembly by Inactivation of Microsomal Triglyceride Transfer Protein Results in a Delayed Removal of apoB-48 from Microsomal and Golgi Membranes of Primary Rat Hepatocytes, J. Lipid Res. 40, 1758–1768.

    PubMed  CAS  Google Scholar 

  26. White, D., Bennett, A., Billett, M., and Salter, A. (1998) The Assembly of Triacylglycerol-Rich Lipoproteins: An Essential Role for the Microsomal Triacylglycerol Transfer Protein, Br. J. Nutr. 80, 219–229.

    PubMed  CAS  Google Scholar 

  27. Levy, E., Stan, S., Delvin, E., Menard, D., Shoulders, C., Garofalo, C., Slight, I., Seidman, E., Mayer, G., and Bendayan, M. (2002) Localization of Microsomal Triglyceride Transfer Protein in the Golgi, J. Biol. Chem. 277, 16470–16477.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, C 3321 Medical Center North, Vanderbilt University School of Medicine, 37232-2561, Nashville, TN

    Klara Valyi-Nagy, Carla Harris & Larry L. Swift

Authors
  1. Klara Valyi-Nagy
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  2. Carla Harris
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  3. Larry L. Swift
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Correspondence to Larry L. Swift.

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Valyi-Nagy, K., Harris, C. & Swift, L.L. The assembly of hepatic very low density lipoproteins: Evidence of a role for the Golgi apparatus. Lipids 37, 879–884 (2002). https://doi.org/10.1007/s11745-002-0974-3

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  • DOI: https://doi.org/10.1007/s11745-002-0974-3

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