Skip to main content
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
. 1980 Jun;77(6):3327–3330. doi: 10.1073/pnas.77.6.3327

A cholesteryl ester transfer complex in human plasma.

P E Fielding, C J Fielding
PMCID: PMC349608  PMID: 6774335

Abstract

Immunoadsorption affinity chromatography has been used to define the structure of lipoproteins in human plasma containing lecithin:cholesterol acyltransferase (EC 2.3.1.43) (LCAT) and transfer protein (apo D). The whole of LCAT was absorbed by antibodies specific for apo D and for apo A-1, indicating that the enzyme is present in plasma exclusively as a complex with its cofactor (apo A-1) and product transfer protein (apo D). About 80% of apo D (but no LCAT) was removed by antibody to apo A-2, indicating the presence of most of apo D in the form of an enzyme-free complex will apo A-1 and apo A-2. After removal of LCAT with antibody to apo D, plasma was unreactive as a substrate with isolated LCAT, but substrate activity was generated by ultracentrifugal flotation with either intact or adsorbed plasma. The apparent stoichiometry of the complex with LCAT (LCAT:apo A-1:apo D) was 1.0:0.9:1.8; that of the complex containing apo A-1, apo A-2, and apo D was 3.9:2.2:1.0.

Full text

PDF
3327

Selected References

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

  1. Albers J. J., Wahl P. W., Cabana V. G., Hazzard W. R., Hoover J. J. Quantitation of apolipoprotein A-I of human plasma high density lipoprotein. Metabolism. 1976 Jun;25(6):633–644. doi: 10.1016/0026-0495(76)90060-3. [DOI] [PubMed] [Google Scholar]
  2. Aron L., Jones S., Fielding C. J. Human plasma lecithin-cholesterol acyltransferase. Characterization of cofactor-dependent phospholipase activity. J Biol Chem. 1978 Oct 25;253(20):7220–7226. [PubMed] [Google Scholar]
  3. Assmann G., Herbert P. N., Fredrickson D. S., Forte T. Isolation and characterization of an abnormal high density lipoprotein in Tangier Diesase. J Clin Invest. 1977 Jul;60(1):242–252. doi: 10.1172/JCI108761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Barter P. J., Conner W. E. The transport of esterified cholesterol in plasma high density lipoproteins of human subjects: a mathematical model. J Lab Clin Med. 1976 Oct;88(4):627–639. [PubMed] [Google Scholar]
  6. Chajek T., Fielding C. J. Isolation and characterization of a human serum cholesteryl ester transfer protein. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3445–3449. doi: 10.1073/pnas.75.7.3445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheung M. C., Albers J. J. The measurement of apolipoprotein A-I and A-II levels in men and women by immunoassay. J Clin Invest. 1977 Jul;60(1):43–50. doi: 10.1172/JCI108767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clifton-Bligh P., Nestel P. J., Whyte H. M. Tangier disease. Report of a case and studies of lipid metabolism. N Engl J Med. 1972 Mar 16;286(11):567–571. doi: 10.1056/NEJM197203162861103. [DOI] [PubMed] [Google Scholar]
  9. Fainaru M., Glangeaud M. C., Eisenberg S. Radioimmunoassay of human high density lipoprotein apo-protein A-1. Biochim Biophys Acta. 1975 Apr 29;386(2):432–443. doi: 10.1016/0005-2795(75)90286-x. [DOI] [PubMed] [Google Scholar]
  10. Fielding C. J., Fielding P. E. Purification and substrate specificity of lecithin-cholesterol acyl transferase from human plasma. FEBS Lett. 1971 Jul 8;15(5):355–358. doi: 10.1016/0014-5793(71)80333-2. [DOI] [PubMed] [Google Scholar]
  11. Fielding C. J. Phospholipid substrate specificity of purified human plasma lecithin:cholesterol acyltransferase. Scand J Clin Lab Invest Suppl. 1974;137:15–17. [PubMed] [Google Scholar]
  12. Fielding C. J., Shore V. G., Fielding P. E. A protein cofactor of lecithin:cholesterol acyltransferase. Biochem Biophys Res Commun. 1972 Feb 25;46(4):1493–1498. doi: 10.1016/0006-291x(72)90776-0. [DOI] [PubMed] [Google Scholar]
  13. Heider J. G., Boyett R. L. The picomole determination of free and total cholesterol in cells in culture. J Lipid Res. 1978 May;19(4):514–518. [PubMed] [Google Scholar]
  14. Kane J. P. A rapid electrophoretic technique for identification of subunit species of apoproteins in serum lipoproteins. Anal Biochem. 1973 Jun;53(2):350–364. doi: 10.1016/0003-2697(73)90081-x. [DOI] [PubMed] [Google Scholar]
  15. LEVY H. B., SOBER H. A. A simple chromatographic method for preparation of gamma globulin. Proc Soc Exp Biol Med. 1960 Jan;103:250–252. doi: 10.3181/00379727-103-25476. [DOI] [PubMed] [Google Scholar]
  16. Osborne J. C., Jr, Brewer H. B., Jr The plasma lipoproteins. Adv Protein Chem. 1977;31:253–337. doi: 10.1016/s0065-3233(08)60220-x. [DOI] [PubMed] [Google Scholar]
  17. Porath J., Axen R., Ernback S. Chemical coupling of proteins to agarose. Nature. 1967 Sep 30;215(5109):1491–1492. doi: 10.1038/2151491a0. [DOI] [PubMed] [Google Scholar]
  18. Shore B., Shore V. Isolation and characterization of polypeptides of human serum lipoproteins. Biochemistry. 1969 Nov;8(11):4510–4516. doi: 10.1021/bi00839a043. [DOI] [PubMed] [Google Scholar]
  19. Stokke K. T., Norum K. R. Determination of lecithin: cholesterol acyltransfer in human blood plasma. Scand J Clin Lab Invest. 1971 Feb;27(1):21–27. doi: 10.3109/00365517109080184. [DOI] [PubMed] [Google Scholar]
  20. Warnick G. R., Albers J. J. A comprehensive evaluation of the heparin-manganese precipitation procedure for estimating high density lipoprotein cholesterol. J Lipid Res. 1978 Jan;19(1):65–76. [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