Cholesterol Efflux from Cells in Culture: Studies with Lipid-Free Acceptors, Reconstituted Particles and Whole Serum

  • G. H. Rothblat
  • P. Yancey
  • W. S. Davidson
  • V. Atger
  • S. Lund-Katz
  • W. J. Johnson
  • M. Llera de la Moya
  • M. C. Phillips
Chapter
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)

Abstract

The movement of cholesterol from cells to acceptor lipoproteins is the first step in the process of reverse cholesterol transport. Both cellular factors and the characteristics of the acceptors modulate the rate at which the cholesterol molecules leave the cell and are picked up by the extracellular acceptors. To gain more information on the factors that influence the efficiency of various acceptors we have conducted a series of studies using acceptor particles of increasing complexity. The simplest system consisted of lipid-free apolipoprotein (apo) Al or synthetic peptides. Increasing complexity was produced when these peptides were reconstituted into disc-like structures composed of phospholipid and the different peptides. The last, and most complex, experimental cholesterol efflux system was one in which whole human serum was added to cells. In all of the studies we have quantitated the release of radiolabeled cholesterol from the cells. In the studies using lipid-free peptides or reconstituted particles the release of the labeled cholesterol reflects the net movement of cholesterol from cells to acceptors, since the acceptors were initially free of cholesterol and no significant cholesterol influx could occur. In the studies using whole serum, the quantitation of labeled cholesterol efflux does not predict the net change in cell cholesterol content since influx would be occurring from a variety of lipoproteins in the serum.

Keywords

Cholesterol Foam Amide Proline Acetyl 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Segrest JP, Jones MK, De Loof H, Brouillette CG, Venkatachalapathi YV, Anantharamaiah GM. The amphipathic helix in the exchangeable apolipoproteins: a review of secondary structure and function. J Lipid Res.1992; 33:141–166. PubMedGoogle Scholar
  2. 2.
    Anantharamaiah GM, Jones JL, Brouillette CG, Schmidt CF, Chung BH, Hughes TA, Bhown AS, Segrest JP. Studies of synthetic peptide analogs of the amphipathic helix. J Biol. Chem.1985;260:10248–10255.PubMedGoogle Scholar
  3. 3.
    Venkatachalapathi YV, Phillips MC, Epand RM, Epand RF, Tytler EM, Segrest JP, Anantharamaiah GM. Effect of end group blockage on the properties of a class A amphipathic helical peptide. Proteins 1993;15:349359.Google Scholar
  4. 4.
    Sokoloff L, Rothblat GH. Regulation of sterol synthesis in L-cells: steady-state and transitional responses. Biochim. Biophys. Acta 1972;280:171181.Google Scholar
  5. 5.
    Castro GR, Fielding CJ. Early incorporation of cell-derived cholesterol into pre-B-migrating high-density lipoprotein. Biochemistry 1988;27:25–29.PubMedCrossRefGoogle Scholar
  6. 6.
    Francone OL, Fielding CJ. Initial steps in reverse cholesterol transport: the role of short-lived cholesterol acceptors. Eur. Heart J. 1990;11:218224.Google Scholar
  7. 7.
    Phillips MC, Johnson WJ, Rothblat GH. Mechanisms and consequences of cellular cholesterol exchange and transfer.. Biochim. Biophys. Acta 1987;906:223–276PubMedCrossRefGoogle Scholar
  8. 8.
    de la Llera Moya M, Atger V, Paul JL, Fournier N, Moatti N, Giral P, Friday KE, Rothblat GH. A cell culture system for screening human serum for ability to promote cellular cholesterol efflux: relationships between serum compnents and efflux, esterification and transfer. Arterioscier. Thromb. 1994;In PressGoogle Scholar
  9. 9.
    Bates SR, Rothblat GH. Regulation of cellular sterol flux and synthesis by human serum lipoproteins. Biochim. Biophys. Acta 1974;360:39–55.Google Scholar
  10. 10.
    Johansson J, Carlson LA, Landou C, Hamsten A. High density lipoproteins and coronary atherosclerosis. A strong inverse relation with the largest particles is confined to normotriglyceridemic patients. Arterioscier. Thromb. 1991;11:174–182.CrossRefGoogle Scholar
  11. 11.
    Lund-Katz S, Hammerschlag B, Phillips MC. Kinetics and mechanism of free cholesterol exchange between human serum high-and low-density lipoprotein. Biochemistry 1982;21:2964–2969.PubMedCrossRefGoogle Scholar
  12. 12.
    Francone OL, Fielding CJ, Fielding PE. Distribution of cell-derived cholesterol among plasma lipoproteins: a comparison of three techniques. J. Lipid Res.1990; 31:2195–2200.PubMedGoogle Scholar
  13. 13.
    Nakamura R, Ohta T, Ikeda Y, Matsuda I. LDL inhibits the mediation of cholesterol efflux from macrophage foam cells by apoA-I-containing lipoproteins. Arterioscier. Thromb. 1993;13:1307–1316.CrossRefGoogle Scholar
  14. 14.
    Johnson WJ, Mahlberg FH, Rothblat GH, Phillips MC. Cholesterol transport between cells and high density lipoproteins. Biochim. Biophys. Acta.1991;1085:273–298.PubMedCrossRefGoogle Scholar
  15. 15.
    Rothblat GH, Mahlberg FH, Johnson WJ, Phillips MC. Apolipoprotein, membrane cholesterol domains, and the regulation of cholesterol efflux. J. Lipid Res.1992;33:1091–1098.PubMedGoogle Scholar
  16. 16.
    Letizia JY, Phillips MC. Effects of apolipoproteins on the kinetics of cholesterol exchange. Biochemistry 1991;30:866–873.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • G. H. Rothblat
    • 1
  • P. Yancey
    • 1
  • W. S. Davidson
    • 1
  • V. Atger
    • 2
  • S. Lund-Katz
    • 1
  • W. J. Johnson
    • 1
  • M. Llera de la Moya
    • 1
  • M. C. Phillips
    • 1
  1. 1.Medical College of PennsylvaniaPhiladelphiaUSA
  2. 2.Hopital BroussaisMedical College of PennsylvaniaParisFrance

Personalised recommendations