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. 1980 Dec;66(6):1351–1362. doi: 10.1172/JCI109988

Radioimmunoassay of human arginine-rich apolipoprotein, apoprotein E. Concentration in blood plasma and lipoproteins as affected by apoprotein E-3 deficiency.

R J Havel, L Kotite, J L Vigne, J P Kane, P Tun, N Phillips, G C Chen
PMCID: PMC371621  PMID: 7440719

Abstract

A radioimmunoassay for apolipoprotein E in human blood serum has been developed that measures equally the major polymorphic species of the protein (apolipoproteins E-1, E-2, E-3, and E-4) and the apo E in the dimer of apolipoproteins E and A-II. The assay is specific and yields values for apolipoprotein E in very low density lipoproteins that agree closely with those obtained by a quantitative electrophoretic method. Apolipoprotein E is also present in at least one species of high density lipoprotein, but the content of apolipoprotein E in the lipoprotein fractions of plasma is uncertain owing to dissociation during ultracentrifugation. The concentration of apolipoprotein E is higher in serum of normolipidemic, premenopausal women than in men of comparable age and is a direct function of the serum triglyceride level. Apolipoprotein E levels are increased out of proportion to triglyceride levels in hyperlipidemic patients with familial dysbetalipoproteinemia (homozygotes for lack of apolipoprotein E-3). Heterozygous relatives of homozygotes have significantly higher apolipoprotein E levels in serum than unaffected relatives. The concentration of partially degraded (remnant) triglyceride-rich lipoproteins also appears to be increased in heterozygotes, who comprise about 15% of the population.

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Selected References

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  1. Brown W. V., Levy R. I., Fredrickson D. S. Studies of the proteins in human plasma very low density lipoproteins. J Biol Chem. 1969 Oct 25;244(20):5687–5694. [PubMed] [Google Scholar]
  2. Curry M. D., McConathy W. J., Alaupovic P., Ledford J. H., Popović M. Determination of human apolipoprotein E by electroimmunoassay. Biochim Biophys Acta. 1976 Aug 9;439(2):413–425. doi: 10.1016/0005-2795(76)90078-7. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Fainaru M., Havel R. J., Imaizumi K. Apoprotein content of plasma lipoproteins of the rat separated by gel chromatography or ultracentrifugation. Biochem Med. 1977 Jun;17(3):347–355. doi: 10.1016/0006-2944(77)90040-0. [DOI] [PubMed] [Google Scholar]
  5. Fainaru M., Havel R. J., Imaizumi K. Radioimmunoassay of arginine-rich apolipoprotein of rat serum. Biochim Biophys Acta. 1977 Jan 25;490(1):144–155. doi: 10.1016/0005-2795(77)90114-3. [DOI] [PubMed] [Google Scholar]
  6. HAVEL R. J., EDER H. A., BRAGDON J. H. The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum. J Clin Invest. 1955 Sep;34(9):1345–1353. doi: 10.1172/JCI103182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Havel R. J., Chao Y., Windler E. E., Kotite L., Guo L. S. Isoprotein specificity in the hepatic uptake of apolipoprotein E and the pathogenesis of familial dysbetalipoproteinemia. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4349–4353. doi: 10.1073/pnas.77.7.4349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Havel R. J., Kane J. P., Kashyap M. L. Interchange of apolipoproteins between chylomicrons and high density lipoproteins during alimentary lipemia in man. J Clin Invest. 1973 Jan;52(1):32–38. doi: 10.1172/JCI107171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Havel R. J., Kane J. P. Primary dysbetalipoproteinemia: predominance of a specific apoprotein species in triglyceride-rich lipoproteins. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2015–2019. doi: 10.1073/pnas.70.7.2015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Havel R. J., Kotite L., Kane J. P. Isoelectric heterogeneity of the cofactor protein for lipoprotein lipase in human blood plasma. Biochem Med. 1979 Feb;21(1):121–138. doi: 10.1016/0006-2944(79)90063-2. [DOI] [PubMed] [Google Scholar]
  11. Huang H., Kauan J. W., Guilbault G. G. Fluorometric enzymatic determination of total cholesterol in serum. Clin Chem. 1975 Oct;21(11):1605–1608. [PubMed] [Google Scholar]
  12. Innerarity T. L., Mahley R. W. Enhanced binding by cultured human fibroblasts of apo-E-containing lipoproteins as compared with low density lipoproteins. Biochemistry. 1978 Apr 18;17(8):1440–1447. doi: 10.1021/bi00601a013. [DOI] [PubMed] [Google Scholar]
  13. Kane J. P., Sata T., Hamilton R. L., Havel R. J. Apoprotein composition of very low density lipoproteins of human serum. J Clin Invest. 1975 Dec;56(6):1622–1634. doi: 10.1172/JCI108245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kashyap M. L., Srivastava L. S., Chen C. Y., Perisutti C. G., Campbell M., Lutmer R. F., Glueck C. J. Radioimmunoassay of human apolipoprotein CII. A study in normal and hypertriglyceridemic subjects. J Clin Invest. 1977 Jul;60(1):171–180. doi: 10.1172/JCI108753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kushwaha R. S., Hazzard W. R., Wahl P. W., Hoover J. J. Type III hyperlipoproteinemia: diagnosis in whole plasma by apolipoprotein-E immunoassay. Ann Intern Med. 1977 Nov;87(5):509–516. doi: 10.7326/0003-4819-87-5-509. [DOI] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Myers L. H., Phillips N. R., Havel R. J. Mathematical evaluation of methods for estimation of the concentration of the major lipid components of human serum lipoproteins. J Lab Clin Med. 1976 Sep;88(3):491–505. [PubMed] [Google Scholar]
  18. Pagnan A., Havel R. J., Kane J. P., Kotite L. Characterization of human very low density lipoproteins containing two electrophoretic populations: double pre-beta lipoproteinemia and primary dysbetalipoproteinemia. J Lipid Res. 1977 Sep;18(5):613–622. [PubMed] [Google Scholar]
  19. Schonfeld G., George P. K., Miller J., Reilly P., Witztum J. Apolipoprotein C-II and C-III levels in hyperlipoproteinemia. Metabolism. 1979 Oct;28(10):1001–1010. doi: 10.1016/0026-0495(79)90004-0. [DOI] [PubMed] [Google Scholar]
  20. Sherrill B. C., Innerarity T. L., Mahley R. W. Rapid hepatic clearance of the canine lipoproteins containing only the E apoprotein by a high affinity receptor. Identity with the chylomicron remnant transport process. J Biol Chem. 1980 Mar 10;255(5):1804–1807. [PubMed] [Google Scholar]
  21. Tada N., Fidge N., Nestel P. Identification and characterization of mixed disulphide complexes of E apoprotein in high density lipoprotein of subjects with acute alcoholic hepatitis. Biochem Biophys Res Commun. 1979 Sep 12;90(1):297–304. doi: 10.1016/0006-291x(79)91624-3. [DOI] [PubMed] [Google Scholar]
  22. Utermann G., Hees M., Steinmetz A. Polymorphism of apolipoprotein E and occurrence of dysbetalipoproteinaemia in man. Nature. 1977 Oct 13;269(5629):604–607. doi: 10.1038/269604a0. [DOI] [PubMed] [Google Scholar]
  23. Utermann G., Jaeschke M., Menzel J. Familial hyperlipoproteinemia type III: deficiency of a specific apolipoprotein (apo E-III) in the very-low-density lipoproteins. FEBS Lett. 1975 Aug 15;56(2):352–355. doi: 10.1016/0014-5793(75)81125-2. [DOI] [PubMed] [Google Scholar]
  24. Utermann G., Pruin N., Steinmetz A. Polymorphism of apolipoprotein E. III. Effect of a single polymorphic gene locus on plasma lipid levels in man. Clin Genet. 1979 Jan;15(1):63–72. [PubMed] [Google Scholar]
  25. Utermann G., Vogelberg K. H., Steinmetz A., Schoenborn W., Pruin N., Jaeschke M., Hees M., Canzler H. Polymorphism of apolipoprotein E. II. Genetics of hyperlipoproteinemia type III. Clin Genet. 1979 Jan;15(1):37–62. [PubMed] [Google Scholar]
  26. Utermann G., Weber W., Beisiegel U. Different mobility in SDS--polyacrylamide gel electrophoresis of apolipoprotein E from phenotypes apo E-N and Apo E-D. FEBS Lett. 1979 May 1;101(1):21–26. [PubMed] [Google Scholar]
  27. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  28. Weisgraber K. H., Mahley R. W. Apoprotein (E--A-II) complex of human plasma lipoproteins. I. Characterization of this mixed disulfide and its identification in a high density lipoprotein subfraction. J Biol Chem. 1978 Sep 10;253(17):6281–6288. [PubMed] [Google Scholar]

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