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. 1970 Dec;49(12):2396–2407. doi: 10.1172/JCI106459

A lipoprotein characterizing obstructive jaundice. II. Isolation and partial characterization of the protein moieties of low density lipoproteins

D Seidel 1,2, P Alaupovic 1,2, R H Furman 1,2, W J McConathy 1,2
PMCID: PMC322741  PMID: 5480863

Abstract

The plasma low density lipoproteins (LDL) in biliary obstruction are characterized almost exclusively by the presence of the immunochemically distinct lipoprotein families, lipoprotein B (LP-B) and lipoprotein X (LP-X). It is suggested that LP-X, with its uniquely high content of unesterified cholesterol and phospholipid, is primarily responsible for the unusual lipid composition of LDL and the abnormal plasma lipid composition in obstructive jaundice.

To show their protein moieties, we isolated LP-X and LP-B from the LDL in plasma obtained from patients with obstructive jaundice. A separation procedure was employed which combines ultracentrifugation, heparin precipitation, and ethanol fractionation. Whereas LP-B was characterized by the presence of apolipoprotein B (ApoB), intact LP-X contained a protein moiety of unique composition consisting of a mixture of albumin (approximately 40%) and the specific apolipoprotein, ApoX (60%). These two protein moieties were separated by preparative ultracentrifugation at d 1.21 g/ml of a solution of partially delipidized LP-X. LP-X thus comprises an albumin-lipoprotein complex in which the masked antigenic site of albumin can be revealed by partial or total delipidization.

Apolipoprotein X, the characteristic nonalbumin protein moiety of intact or partially delipidized LP-X, was immunochemically different from ApoA, ApoB, albumin, γ-globulins, and other serum proteins. The results of analytical ultracentrifugation and the immunochemical and electrophoretic properties of ApoX indicated it to be a complex protein consisting possibly of several nonidentical polypeptides. ApoX was characterized by its amino acid composition, and by serine and threonine as the major N-terminal and alanine as the major C-terminal amino acids. It has been suggested that ApoX is similar to, if not identical with, apolipoprotein C.

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Selected References

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

  1. AYRAULT-JARRIER M., LEVY G., WALD R., POLONOVSKI J. [Separation by ultracentrifugation of the alpha-lipoproteins of normal human serum]. Bull Soc Chim Biol (Paris) 1963;45:349–359. [PubMed] [Google Scholar]
  2. Alaupovic P., Furman R. H., Falor W. H., Sullivan M. L., Walraven S. L., Olson A. C. Isolation and characterization of human chyle chylomicrons and lipoproteins. Ann N Y Acad Sci. 1968 Nov 21;149(2):791–807. doi: 10.1111/j.1749-6632.1968.tb53836.x. [DOI] [PubMed] [Google Scholar]
  3. Alaupovic P., Furman R. H., Falor W. H., Sullivan M. L., Walraven S. L., Olson A. C. Isolation and characterization of human chyle chylomicrons and lipoproteins. Ann N Y Acad Sci. 1968 Nov 21;149(2):791–807. doi: 10.1111/j.1749-6632.1968.tb53836.x. [DOI] [PubMed] [Google Scholar]
  4. Alaupovic P., Seidel D., McConathy W. J., Furman R. H. Identification of the protein moiety of an abnormal human plasma low-density lipoprotein in obstructive jaundice. FEBS Lett. 1969 Jul;4(2):113–116. doi: 10.1016/0014-5793(69)80209-7. [DOI] [PubMed] [Google Scholar]
  5. BENSON J. V., Jr, PATTERSON J. A. ACCELERATED AUTOMATIC CHROMATOGRAPHIC ANALYSIS OF AMINO ACIDS ON A SPHERICAL RESIN. Anal Chem. 1965 Aug;37:1108–1110. doi: 10.1021/ac60228a008. [DOI] [PubMed] [Google Scholar]
  6. Beaumont J. L., Jacotot B., Beaumont V., Warnet J., Vilain C. Myélome, hyperlipidémie et xanthomatose. Nouv Rev Fr Hematol. 1965 May-Jun;5(3):507–517. [PubMed] [Google Scholar]
  7. Brown W. V., Levy R. I., Fredrickson D. S. Further separation of the apoproteins of the human plasma very low density lipoproteins. Biochim Biophys Acta. 1970 Mar 31;200(3):573–575. doi: 10.1016/0005-2795(70)90115-7. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Burstein M., Caroli J. Isolement et étude des lipoprotéines sériques anormales au cours des ictères par rétention après floculation par le polyvinyl-pyrrolidone. Rev Fr Etud Clin Biol. 1968 Apr;13(4):387–391. [PubMed] [Google Scholar]
  10. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  11. DELALLA L., LEVINE L., BROWN R. K. Immunologic studies of human high density lipoproteins. J Exp Med. 1957 Aug 1;106(2):261–271. doi: 10.1084/jem.106.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. EDER H. A., RUSS E. M., PRITCHETT R. A. R., WILBER M. M., BARR D. P. Protein-lipid relationships in human plasma: in biliary cirrhosis, obstructive jaundice, and acute hepatitis. J Clin Invest. 1955 Jul;34(7 Pt 1):1147–1162. doi: 10.1172/JCI103163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. FURMAN R. H., CONRAD L. L. Ultracentrifugal characterization of the lipoprotein spectrum in obstructive jaundice: studies of serum lipid relationships in intra-and extrahepatic biliary obstruction. J Clin Invest. 1957 May;36(5):713–722. doi: 10.1172/JCI103474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fredrickson D. S., Levy R. I., Lees R. S. Fat transport in lipoproteins--an integrated approach to mechanisms and disorders. N Engl J Med. 1967 Feb 2;276(5):273–concl. doi: 10.1056/NEJM196702022760507. [DOI] [PubMed] [Google Scholar]
  15. GRABAR P., WILLIAMS C. A., Jr Méthode immuno-électrophorétique d'analyse de mélanges de substances antigéniques. Biochim Biophys Acta. 1955 May;17(1):67–74. doi: 10.1016/0006-3002(55)90320-6. [DOI] [PubMed] [Google Scholar]
  16. GUSTAFSON A., ALAUPOVIC P., FURMAN R. H. STUDIES OF THE COMPOSITION AND STRUCTURE OF SERUM LIPOPROTEINS: ISOLATION, PURIFICATION, AND CHARACTERIZATION OF VERY LOW DENSITY LIPOPROTEINS OF HUMAN SERUM. Biochemistry. 1965 Mar;4:596–605. doi: 10.1021/bi00879a033. [DOI] [PubMed] [Google Scholar]
  17. GUSTAFSON A., ALAUPOVIC P., FURMAN R. H. STUDIES OF THE COMPOSITION AND STRUCTURE OF SERUM LIPOPROTEINS: PHYSICAL-CHEMICAL CHARACTERIZATION OF PHOSPHOLIPID-PROTEIN RESIDUES OBTAINED FROM VERY-LOW-DENSITY HUMAN SERUM LIPOPROTEINS. Biochim Biophys Acta. 1964 Dec 2;84:767–769. doi: 10.1016/0926-6542(64)90040-x. [DOI] [PubMed] [Google Scholar]
  18. Granda J. L., Scanu A. Solubilization and properties of the apoproteins of the very low- and low-density lipoproteins of human serum. Biochemistry. 1966 Oct;5(10):3301–3308. doi: 10.1021/bi00874a032. [DOI] [PubMed] [Google Scholar]
  19. Gros C., Labouesse B. Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem. 1969 Feb;7(4):463–470. doi: 10.1111/j.1432-1033.1969.tb19632.x. [DOI] [PubMed] [Google Scholar]
  20. Gustafson A., Alaupovic P., Furman R. H. Studies of the composition and structure of serum lipoproteins. Separation and characterization of phospholipid-protein residues obtained by partial delipidization of very low density lipoproteins of human serum. Biochemistry. 1966 Feb;5(2):632–640. doi: 10.1021/bi00866a033. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. HIRS C. H. The oxidation of ribonuclease with performic acid. J Biol Chem. 1956 Apr;219(2):611–621. [PubMed] [Google Scholar]
  23. LEWIS L. A., PAGE I. H. AN UNUSUAL SERUM LIPOPROTEIN-GLOBULIN COMPLEX IN A PATIENT WITH HYPERLIPEMIA. Am J Med. 1965 Feb;38:286–297. doi: 10.1016/0002-9343(65)90183-x. [DOI] [PubMed] [Google Scholar]
  24. Lee D. M., Alaupovic P. Studies of the composition and structure of plasma lipoproteins. Isolation, composition, and immunochemical characterization of low density lipoprotein subfractions of human plasma. Biochemistry. 1970 May 26;9(11):2244–2252. doi: 10.1021/bi00813a004. [DOI] [PubMed] [Google Scholar]
  25. Lemaire A., Etienne G., Etienne J., Polonovski J., Housset E., Cottet J. Un vouveau test biologique de la cholostase. Presse Med. 1965 Dec 25;73(56):3245–3247. [PubMed] [Google Scholar]
  26. Levy R. I., Lees R. S., Fredrickson D. S. The nature of pre beta (very low density) lipoproteins. J Clin Invest. 1966 Jan;45(1):63–77. doi: 10.1172/JCI105324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Levy R. S., Lynch A. C., McGee E. D., Mehl J. W. Amino acid composition of the proteins from chylomicrons and human serum lipoproteins. J Lipid Res. 1967 Sep;8(5):463–472. [PubMed] [Google Scholar]
  28. Lewis L. A., Van Ommen R. A., Page I. H. Association of cold-precipitability with beta-lipoprotein and cryoglobulin. Am J Med. 1966 May;40(5):785–793. doi: 10.1016/0002-9343(66)90161-6. [DOI] [PubMed] [Google Scholar]
  29. Margolis S., Langdon R. G. Studies on human serum beta-1-lipoprotein. I. Amino acid composition. J Biol Chem. 1966 Jan 25;241(2):469–476. [PubMed] [Google Scholar]
  30. POULIK M. D. Starch gel electrophoresis in a discontinous system of buffers. Nature. 1957 Dec 28;180(4600):1477–1479. doi: 10.1038/1801477a0. [DOI] [PubMed] [Google Scholar]
  31. RUSS E. M., RAYMUNT J., BARR D. P. Lipoproteins in primary biliary cirrhosis. J Clin Invest. 1956 Feb;35(2):133–144. doi: 10.1172/JCI103257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rudman D., Garcia L. A., Abell L. L., Akgun S. Observations on the protein components of human plasma high- and low-density lipoproteins. Biochemistry. 1968 Sep;7(9):3136–3148. doi: 10.1021/bi00849a017. [DOI] [PubMed] [Google Scholar]
  33. SCANU A., LEWIS L. A., BUMPUS F. M. Separation and characterization of the protein moiety of human alpha1-lipoprotein. Arch Biochem Biophys. 1958 Apr;74(2):390–397. doi: 10.1016/0003-9861(58)90009-2. [DOI] [PubMed] [Google Scholar]
  34. SCANU A., LEWIS L. A., PAGE I. H. Studies on the antigenicity of beta- and alpha-lipoproteins of human serum. J Exp Med. 1958 Aug 1;108(2):185–196. doi: 10.1084/jem.108.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. SMITHIES O. Zone electrophoresis in starch gels: group variations in the serum proteins of normal human adults. Biochem J. 1955 Dec;61(4):629–641. doi: 10.1042/bj0610629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scanu A., Granda J. L. Effects of ultracentrifugation on the human serum high-density (1.063 less than p less than 1.21 g/ml) lipoprotein. Biochemistry. 1966 Feb;5(2):446–455. doi: 10.1021/bi00866a008. [DOI] [PubMed] [Google Scholar]
  37. Seidel D., Alaupovic P., Furman R. H. A lipoprotein characterizing obstructive jaundice. I. Method for quantitative separation and identification of lipoproteins in jaundiced subjects. J Clin Invest. 1969 Jul;48(7):1211–1223. doi: 10.1172/JCI106085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shore B., Shore V. Heterogeneity in protein subunits of human serum high-density lipoproteins. Biochemistry. 1968 Aug;7(8):2773–2777. doi: 10.1021/bi00848a011. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Shore V., Shore B. Some physical and chemical studies on the protein moiety of a high-density (1.126-1.195 g-ml) lipoprotein fraction of human serum. Biochemistry. 1967 Jul;6(7):1962–1969. doi: 10.1021/bi00859a012. [DOI] [PubMed] [Google Scholar]
  41. Shore V., Shore B. Some physical and chemical studies on two polypeptide components of high-density lipoproteins of human serum. Biochemistry. 1968 Oct;7(10):3396–3403. doi: 10.1021/bi00850a013. [DOI] [PubMed] [Google Scholar]
  42. Skipski V. P., Barclay M., Barclay R. K., Fetzer V. A., Good J. J., Archibald F. M. Lipid composition of human serum lipoproteins. Biochem J. 1967 Aug;104(2):340–352. doi: 10.1042/bj1040340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Switzer S. Plasma lipoproteins in liver disease. I. Immunologically distinct low-density lipoproteins in patients with biliary obstruction. J Clin Invest. 1967 Nov;46(11):1855–1866. doi: 10.1172/JCI105676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. THOMPSON E. O. The N-terminal sequence of serum albumins; observations on the thiohydantoin method. J Biol Chem. 1954 Jun;208(2):565–572. [PubMed] [Google Scholar]
  45. Van Orden D. E., Treffers H. P. A demonstration of the coprecipitation of beta-lipoproteins with specific precipitates of chicken antibodies and human serum albumin. J Immunol. 1968 Mar;100(3):664–674. [PubMed] [Google Scholar]
  46. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]

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