Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Crystal structure of cholesteryl ester transfer protein reveals a long tunnel and four bound lipid molecules

Nature Structural & Molecular Biology volume 14, pages 106–113 (2007)Cite this article

Abstract

Cholesteryl ester transfer protein (CETP) shuttles various lipids between lipoproteins, resulting in the net transfer of cholesteryl esters from atheroprotective, high-density lipoproteins (HDL) to atherogenic, lower-density species. Inhibition of CETP raises HDL cholesterol and may potentially be used to treat cardiovascular disease. Here we describe the structure of CETP at 2.2-Å resolution, revealing a 60-Å-long tunnel filled with two hydrophobic cholesteryl esters and plugged by an amphiphilic phosphatidylcholine at each end. The two tunnel openings are large enough to allow lipid access, which is aided by a flexible helix and possibly also by a mobile flap. The curvature of the concave surface of CETP matches the radius of curvature of HDL particles, and potential conformational changes may occur to accommodate larger lipoprotein particles. Point mutations blocking the middle of the tunnel abolish lipid-transfer activities, suggesting that neutral lipids pass through this continuous tunnel.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Overall structure of CETP.
Figure 2: Bound lipids and the tunnel.
Figure 3: Stereo view of the lipid-binding sites.
Figure 4: The mobile helix X and the Ω1 flap.
Figure 5: Concave surface and HDL binding.
Figure 6: Proposed mechanism for CETP-mediated heteroexchange.

Similar content being viewed by others

Accession codes

Primary accessions

Protein Data Bank

References

  1. National Center for Chronic Disease Prevention and Health Promotion. Preventing heart disease and stroke. Centers for Disease Control and Prevention http://www.cdc.gov/nccdphp/publications/factsheets/Prevention/cvh.htm (2005).

  2. Gordon, D.J. et al. High-density lipoprotein cholesterol and cardiovascular disease. Four prospective American studies. Circulation 79, 8–15 (1989).

    Article  CAS  Google Scholar 

  3. Boden, W.E. High-density lipoprotein cholesterol as an independent risk factor in cardiovascular disease: assessing the data from Framingham to the Veterans Affairs High-Density Lipoprotein Intervention Trial. Am. J. Cardiol. 86, 19L–22L (2000).

    Article  CAS  Google Scholar 

  4. Rubins, H.B. et al. Gemfibrozil for the secondary prevention of coronary heart disease in men with low levels of high-density lipoprotein cholesterol. Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial Study Group. N. Engl. J. Med. 341, 410–418 (1999).

    Article  CAS  Google Scholar 

  5. Shepherd, J., Betteridge, J., Van Gaal, L. & European Consensus Panel. Nicotinic acid in the management of dyslipidaemia associated with diabetes and metabolic syndrome: a position paper developed by a European Consensus Panel. Curr. Med. Res. Opin. 21, 665–682 (2005).

    Article  CAS  Google Scholar 

  6. Okamoto, H. et al. A cholesteryl ester transfer protein inhibitor attenuates atherosclerosis in rabbits. Nature 406, 203–207 (2000).

    Article  CAS  Google Scholar 

  7. Clark, R.W. et al. Raising high-density lipoprotein in humans through inhibition of cholesteryl ester transfer protein: an initial multidose study of torcetrapib. Arterioscler. Thromb. Vasc. Biol. 24, 490–497 (2004).

    Article  CAS  Google Scholar 

  8. Brousseau, M.E. et al. Effects of an inhibitor of cholesteryl ester transfer protein on HDL cholesterol. N. Engl. J. Med. 350, 1505–1515 (2004).

    Article  CAS  Google Scholar 

  9. Linsel-Nitschke, P. & Tall, A.R. HDL as a target in the treatment of atherosclerotic cardiovascular disease. Nat. Rev. Drug Discov. 4, 193–205 (2005).

    Article  CAS  Google Scholar 

  10. Tall, A.R. Plasma cholesteryl ester transfer protein. J. Lipid Res. 34, 1255–1274 (1993).

    CAS  PubMed  Google Scholar 

  11. Barter, P.J. et al. Cholesteryl ester transfer protein: a novel target for raising HDL and inhibiting atherosclerosis. Arterioscler. Thromb. Vasc. Biol. 23, 160–167 (2003).

    Article  CAS  Google Scholar 

  12. Kawano, K., Qin, S.C., Lin, M., Tall, A.R. & Jiang, X.C. Cholesteryl ester transfer protein and phospholipid transfer protein have nonoverlapping functions in vivo. J. Biol. Chem. 275, 29477–29481 (2000).

    Article  CAS  Google Scholar 

  13. Klerkx, A.H. et al. Cholesteryl ester transfer protein (CETP) inhibition: beyond raising high-density lipoprotein cholesterol levels: pathways by which modulation of CETP may alter atherogenesis. Arterioscler. Thromb. Vasc. Biol. 26, 706–715 (2006).

    Article  CAS  Google Scholar 

  14. Mahley, R.W., Huang, Y. & Weisgraber, K.H. Putting cholesterol in its place: apoE and reverse cholesterol transport. J. Clin. Invest. 116, 1226–1229 (2006).

    Article  CAS  Google Scholar 

  15. Bruce, C., Beamer, L.J. & Tall, A.R. The implications of the structure of the bactericidal/permeability-increasing protein on the lipid-transfer function of the cholesteryl ester transfer protein. Curr. Opin. Struct. Biol. 8, 426–434 (1998).

    Article  CAS  Google Scholar 

  16. Beamer, L.J. Structure of human BPI (bactericidal/permeability-increasing protein) and implications for related proteins. Biochem. Soc. Trans. 31, 791–794 (2003).

    Article  CAS  Google Scholar 

  17. Beamer, L.J., Carroll, S.F. & Eisenberg, D. Crystal structure of human BPI and two bound phospholipids at 2.4 angstrom resolution. Science 276, 1861–1864 (1997).

    Article  CAS  Google Scholar 

  18. Hamilton, J.A. Fatty acid interactions with proteins: what X-ray crystal and NMR solution structures tell us. Prog. Lipid Res. 43, 177–199 (2004).

    Article  CAS  Google Scholar 

  19. Alpy, F. & Tomasetto, C. Give lipids a START: the StAR-related lipid transfer (START) domain in mammals. J. Cell Sci. 118, 2791–2801 (2005).

    Article  CAS  Google Scholar 

  20. Malinina, L., Malakhova, M.L., Teplov, A., Brown, R.E. & Patel, D.J. Structural basis for glycosphingolipid transfer specificity. Nature 430, 1048–1053 (2004).

    Article  CAS  Google Scholar 

  21. Im, Y.J., Raychaudhuri, S., Prinz, W.A. & Hurley, J.H. Structural mechanism for sterol sensing and transport by OSBP-related proteins. Nature 437, 154–158 (2005).

    Article  CAS  Google Scholar 

  22. Sha, B., Phillips, S.E., Bankaitis, V.A. & Luo, M. Crystal structure of the Saccharomyces cerevisiae phosphatidylinositol-transfer protein. Nature 391, 506–510 (1998).

    Article  CAS  Google Scholar 

  23. Qiu, X. & Janson, C.A. Structure of apo acyl carrier protein and a proposal to engineer protein crystallization through metal ions. Acta Crystallogr. D Biol. Crystallogr. 60, 1545–1554 (2004).

    Article  Google Scholar 

  24. Connolly, D.T. et al. Physical and kinetic characterization of recombinant human cholesteryl ester transfer protein. Biochem. J. 320, 39–47 (1996).

    Article  CAS  Google Scholar 

  25. Clark, R.W., Ruggeri, R.B., Cunningham, D. & Bamberger, M.J. Description of the torcetrapib series of cholesteryl ester transfer protein inhibitors, including mechanism of action. J. Lipid Res. 47, 537–552 (2006).

    Article  CAS  Google Scholar 

  26. Wright, C.S., Li, S.C. & Rastinejad, F. Crystal structure of human GM2-activator protein with a novel β-cup topology. J. Mol. Biol. 304, 411–422 (2000).

    Article  CAS  Google Scholar 

  27. Wang, S., Kussie, P., Deng, L. & Tall, A. Defective binding of neutral lipids by a carboxyl-terminal deletion mutant of cholesteryl ester transfer protein. Evidence for a carboxyl-terminal cholesteryl ester binding site essential for neutral lipid transfer activity. J. Biol. Chem. 270, 612–618 (1995).

    Article  CAS  Google Scholar 

  28. Rajaram, O.V. & Sawyer, W.H. Penetration of an emulsion surface by cholesteryl transfer protein. Eur. Biophys. J. 25, 31–36 (1996).

    Article  CAS  Google Scholar 

  29. Zheng, K.Q. et al. A novel missense mutation (L296Q) in cholesteryl ester transfer protein gene related to coronary heart disease. Acta Biochim. Biophys. Sin. (Shanghai) 36, 33–36 (2004).

    Article  CAS  Google Scholar 

  30. Bruce, C. et al. Molecular determinants of plasma cholesteryl ester transfer protein binding to high density lipoproteins. J. Biol. Chem. 270, 11532–11542 (1995).

    Article  CAS  Google Scholar 

  31. Jiang, X.C. et al. Point mutagenesis of positively charged amino acids of cholesteryl ester transfer protein: conserved residues within the lipid transfer/lipopolysaccharide binding protein gene family essential for function. Biochemistry 34, 7258–7263 (1995).

    Article  CAS  Google Scholar 

  32. Ohnishi, T., Oikawa, K., Kay, C.M. & Yokoyama, S. Modulation of substrate selectivity in plasma lipid transfer protein reaction over structural variation of lipid particle. Biochim. Biophys. Acta 1254, 117–126 (1995).

    Article  Google Scholar 

  33. Serdyuk, A.P. & Morton, R.E. Lipid transfer inhibitor protein defines the participation of lipoproteins in lipid transfer reactions: CETP has no preference for cholesteryl esters in HDL versus LDL. Arterioscler. Thromb. Vasc. Biol. 19, 718–726 (1999).

    Article  CAS  Google Scholar 

  34. Peter, B.J. et al. BAR domains as sensors of membrane curvature: the amphiphysin BAR structure. Science 303, 495–499 (2004).

    Article  CAS  Google Scholar 

  35. Miller, K.W. & Small, D.M. Surface-to-core and interparticle equilibrium distributions of triglyceride-rich lipoprotein lipids. J. Biol. Chem. 258, 13772–13784 (1983).

    CAS  PubMed  Google Scholar 

  36. Hamilton, J.A. & Small, D.M. Solubilization and localization of cholesteryl oleate in egg phosphatidylcholine vesicles. A carbon 13 NMR study. J. Biol. Chem. 257, 7318–7321 (1982).

    CAS  PubMed  Google Scholar 

  37. Hamilton, J.A., Miller, K.W. & Small, D.M. Solubilization of triolein and cholesteryl oleate in egg phosphatidylcholine vesicles. J. Biol. Chem. 258, 12821–12826 (1983).

    CAS  Google Scholar 

  38. Morton, R.E. & Steinbrunner, J.V. Concentration of neutral lipids in the phospholipid surface of substrate particles determines lipid transfer protein activity. J. Lipid Res. 31, 1559–1567 (1990).

    CAS  PubMed  Google Scholar 

  39. Morton, R.E. & Zilversmit, D.B. Inter-relationship of lipids transferred by the lipid-transfer protein isolated from human lipoprotein-deficient plasma. J. Biol. Chem. 258, 11751–11757 (1983).

    CAS  PubMed  Google Scholar 

  40. Ko, K.W., Ohnishi, T. & Yokoyama, S. Triglyceride transfer is required for net cholesteryl ester transfer between lipoproteins in plasma by lipid transfer protein. Evidence for a hetero-exchange transfer mechanism demonstrated by using novel monoclonal antibodies. J. Biol. Chem. 269, 28206–28213 (1994).

    CAS  PubMed  Google Scholar 

  41. Urlaub, G., Kas, E., Carothers, A.M. & Chasin, L.A. Deletion of the diploid dihydrofolate reductase locus from cultured mammalian cells. Cell 33, 405–412 (1983).

    Article  CAS  Google Scholar 

  42. Collaborative Computational Project, Number 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  43. Lloyd, D.B. et al. Cholesteryl ester transfer protein variants have differential stability but uniform inhibition by torcetrapib. J. Biol. Chem. 280, 14918–14922 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank the staffs in the X-ray group and synchrotrons for assistance in data collection, G. Andrews, M. Bamberger, L. Morehouse, D. Perry, R. Ruggeri, K. Ranney, I. Reininger, M. Tu and P. Zagouras for insightful discussions and S. Liu, J. Boyd, D. Cunningham, T. Dickinson, J. Duerr, B. King, T. Lanzetti, W. Lin, P. Loulakis, M. Mansour, A. McColl, T. McLellan, F. Rajamohan, M. Rosner, M. Tardie and Z. Xie for supporting work.

Author information

Authors and Affiliations

  1. Pfizer Global Research and Development, Eastern Point Road, Groton, 06430, Connecticut, USA

    Xiayang Qiu, Anil Mistry, Mark J Ammirati, Boris A Chrunyk, Ronald W Clark, Yang Cong, Jeffrey S Culp, Dennis E Danley, Thomas B Freeman, Kieran F Geoghegan, Matthew C Griffor, Steven J Hawrylik, Cheryl M Hayward, Preston Hensley, Lise R Hoth, George A Karam, Maruja E Lira, David B Lloyd, Katherine M McGrath, Kim J Stutzman-Engwall, Ann K Subashi, Timothy A Subashi, John F Thompson, Ing-Kae Wang, Honglei Zhao & Andrew P Seddon

Authors
  1. Xiayang Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anil Mistry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mark J Ammirati
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Boris A Chrunyk
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ronald W Clark
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yang Cong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jeffrey S Culp
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dennis E Danley
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Thomas B Freeman
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Kieran F Geoghegan
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Matthew C Griffor
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Steven J Hawrylik
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Cheryl M Hayward
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Preston Hensley
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Lise R Hoth
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. George A Karam
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Maruja E Lira
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. David B Lloyd
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Katherine M McGrath
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Kim J Stutzman-Engwall
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Ann K Subashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  22. Timothy A Subashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  23. John F Thompson
    View author publications

    You can also search for this author in PubMed Google Scholar

  24. Ing-Kae Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  25. Honglei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  26. Andrew P Seddon
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

M.C.G., S.J.H., T.A.S., I.-K.W., H.Z., K.M.M., K.J.S.-E., T.B.F., L.R.H., K.F.G., Y.C., G.A.K., B.A.C., J.S.C. and A.K.S. provided key reagents. A.M., M.J.A. and D.E.D. crystallized the proteins. X.Q. solved the structure. M.E.L., D.B.L., J.F.T. and R.W.C. contributed to mutagenesis and assays. X.Q., P.H., C.M.H. and A.P.S coordinated the research and data review. X.Q., J.F.T., K.F.G. and A.P.S. wrote the paper.

Corresponding author

Correspondence to Xiayang Qiu.

Ethics declarations

Competing interests

The authors are employees of Pfizer Inc., which develops and markets medicines to treat cardiovascular disease.

Supplementary information

Supplementary Fig. 1

Structure-based alignment of human CETP species and BPI. (PDF 436 kb)

Supplementary Data

Additional methods and data. (PDF 165 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Qiu, X., Mistry, A., Ammirati, M. et al. Crystal structure of cholesteryl ester transfer protein reveals a long tunnel and four bound lipid molecules. Nat Struct Mol Biol 14, 106–113 (2007). https://doi.org/10.1038/nsmb1197

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nsmb1197

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing