Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1982 Jun 1;203(3):727–734. doi: 10.1042/bj2030727

The characterization of lipoprotein lipase isolated from the post-heparin plasma of the rainbow trout, Salmo gairdneri Richardson.

E R Skinner, A M Youssef
PMCID: PMC1158289  PMID: 7115310

Abstract

1. Intravenous injection of heparin into the trout resulted in the appearance in the plasma of a lipase with the properties of lipoprotein lipase. 2. The enzyme was purified to apparent electrophoretic homogeneity by means of heparin-Sepharose affinity chromatography. The enzyme was eluted with 1.5 M-NaCl and had a specific activity approx. 450-fold that of the post-heparin plasma. 3. The activity of the purified enzyme was inhibited by 1.0 M-NaCl and protamine sulphate and was stimulated between 3- and 8.8-fold by the addition of trout plasma. 4. The activity was strongly stimulated by trout very low density lipoproteins and to a lesser extent by high density lipoproteins. 5. The isolated enzyme fraction gave a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and had an apparent subunit M4 of 63 000. 6. These results suggest that the uptake of lipid by the tissues in the trout can occur by a process similar to that in mammals.

Full text

PDF
727

Images in this article

730Fig. 3.
on p.730
731Fig. 6.
on p.731

Selected References

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

  1. Augustin J., Freeze H., Tejada P., Brown W. V. A comparison of molecular properties of hepatic triglyceride lipase and lipoprotein lipase from human post-heparin plasma. J Biol Chem. 1978 May 10;253(9):2912–2920. [PubMed] [Google Scholar]
  2. Becht I., Schrecker O., Klose G., Greten H. Purification of human plasma lipoprotein lipase. Biochim Biophys Acta. 1980 Dec 5;620(3):583–591. doi: 10.1016/0005-2760(80)90150-2. [DOI] [PubMed] [Google Scholar]
  3. Belfrage P., Vaughan M. Simple liquid-liquid partition system for isolation of labeled oleic acid from mixtures with glycerides. J Lipid Res. 1969 May;10(3):341–344. [PubMed] [Google Scholar]
  4. Bensadoun A., Ehnholm C., Steinberg D., Brown W. V. Purification and characterization of lipoprotein lipase from pig adipose tissue. J Biol Chem. 1974 Apr 10;249(7):2220–2227. [PubMed] [Google Scholar]
  5. Chapman M. J., Goldstein S., Mills G. L., Leger C. Distribution and characterization of the serum lipoproteins and their apoproteins in the rainbow trout (Salmo gairdnerii). Biochemistry. 1978 Oct 17;17(21):4455–4464. doi: 10.1021/bi00614a015. [DOI] [PubMed] [Google Scholar]
  6. Chung J., Scanu A. M. Isolation, molecular properties, and kinetic characterization of lipoprotein lipase from rat heart. J Biol Chem. 1977 Jun 25;252(12):4202–4209. [PubMed] [Google Scholar]
  7. 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]
  8. Egelrud T., Olivecrona T. The purification of a lipoprotein lipase from bovine skim milk. J Biol Chem. 1972 Oct 10;247(19):6212–6217. [PubMed] [Google Scholar]
  9. Eisenberg S., Levy R. I. Lipoprotein metabolism. Adv Lipid Res. 1975;13:1–89. [PubMed] [Google Scholar]
  10. Fielding C. J. Human lipoprotein lipase. II. Inhibition of enzyme activity by plasma low density lipoproteins. Biochim Biophys Acta. 1970 Apr 22;206(1):118–124. doi: 10.1016/0005-2744(70)90088-4. [DOI] [PubMed] [Google Scholar]
  11. Hernell O., Egelrud T., Olivecrona T. Serum-stimulated lipases (lipoprotein lipases). Immunological crossreaction between the bovine and the human enzymes. Biochim Biophys Acta. 1975 Feb 13;381(2):233–241. [PubMed] [Google Scholar]
  12. Hernell O., Olivecrona T. Human milk lipases. I. Serum-stimulated lipase. J Lipid Res. 1974 Jul;15(4):367–374. [PubMed] [Google Scholar]
  13. Iverius P. H., Ostlund-Lindqvist A. M. Lipoprotein lipase from bovine milk. Isolation procedure, chemical characterization, and molecular weight analysis. J Biol Chem. 1976 Dec 25;251(24):7791–7795. [PubMed] [Google Scholar]
  14. Kompiang I. P., Bensadoun A., Yang M. W. Effect of an anti-lipoprotein lipase serum on plasma triglyceride removal. J Lipid Res. 1976 Sep;17(5):498–505. [PubMed] [Google Scholar]
  15. Kuusi T., Schröder T., Bång B., Lempinen M., Ehnholm C. Selective measurement of triacylglycerol lipase activities in pig post-heparin plasma. Biochim Biophys Acta. 1979 Jun 21;573(3):443–450. doi: 10.1016/0005-2760(79)90219-4. [DOI] [PubMed] [Google Scholar]
  16. LIS E. W., TINOCO J., OKEY R. A micromethod for fractionation of lipids by silicic acid chromatography. Anal Biochem. 1961 Apr;2:100–106. doi: 10.1016/0003-2697(61)90058-6. [DOI] [PubMed] [Google Scholar]
  17. LaRosa J. C., Levy R. I., Herbert P., Lux S. E., Fredrickson D. S. A specific apoprotein activator for lipoprotein lipase. Biochem Biophys Res Commun. 1970 Oct 9;41(1):57–62. doi: 10.1016/0006-291x(70)90468-7. [DOI] [PubMed] [Google Scholar]
  18. LaRosa J. C., Levy R. I., Windmueller H. G., Fredrickson D. S. Comparison of the triglyceride lipase of liver, adipose tissue, and postheparin plasma. J Lipid Res. 1972 May;13(3):356–363. [PubMed] [Google Scholar]
  19. March S. C., Parikh I., Cuatrecasas P. A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal Biochem. 1974 Jul;60(1):149–152. doi: 10.1016/0003-2697(74)90139-0. [DOI] [PubMed] [Google Scholar]
  20. Ostlung-Lindqvist A. M., Boberg J. Purification of salt resistant lipase and lipoprotein lipase from human post-heparin plasma. FEBS Lett. 1977 Nov 15;83(2):231–236. doi: 10.1016/0014-5793(77)81011-9. [DOI] [PubMed] [Google Scholar]
  21. ROBINSON D. S. CHANGES IN THE LIPOLYTIC ACTIVITY OF THE GUINEA PIG MAMMARY GLAND AT PARTURITION. J Lipid Res. 1963 Jan;4:21–23. [PubMed] [Google Scholar]
  22. Robinson J. S., Mead J. F. Lipid absorption and deposition in rainbow trout (Salmo gairdnerii). Can J Biochem. 1973 Jul;51(7):1050–1058. doi: 10.1139/o73-137. [DOI] [PubMed] [Google Scholar]
  23. Rogie A., Skinner E. R. Some aspects of lipid transport in the rainbow trout. Biochem Soc Trans. 1981 Feb;9(1):59–60. doi: 10.1042/bst0090059. [DOI] [PubMed] [Google Scholar]
  24. Skinner E. R., Rogie A. The isolation and partial characterization of the serum lipoproteins and apolipoproteins of the rainbow trout. Biochem J. 1978 Aug 1;173(2):507–520. doi: 10.1042/bj1730507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Skinner E. R., Youssef A. M., Plack P. A. Lipoprotein lipase activity in the post-heparin plasma and adipose tissue of the rainbow trout (Salmo gairdnerii) [proceedings]. Biochem Soc Trans. 1980 Feb;8(1):74–74. doi: 10.1042/bst0080074. [DOI] [PubMed] [Google Scholar]
  26. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES