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. 1985 May 15;228(1):137–146. doi: 10.1042/bj2280137

Degradation of endocytosed insulin in rat liver is mediated by low-density vesicles.

R J Pease, G D Smith, T J Peters
PMCID: PMC1144962  PMID: 3890838

Abstract

Uptake of 125I-insulin by the liver of intact rats is followed by rapid translocation of label to low-density vesicles. Subcellular-fractionation studies indicate that, although the 125I associated with these vesicles is predominantly trichloroacetic acid-precipitable, there is an acid-soluble component arising from processing of the hormone in vivo. H.p.l.c. analysis indicates that the acid-precipitable 125I is not intact iodoinsulin, but may correspond to 'clipped insulin'. Isolated low-density vesicles degrade the acid-precipitable iodopeptide intravesicularly when incubated at 37 degrees C in iso-osmotic medium at pH7. The rate constant for intravesicular degradation is consistent with the rate of insulin clearance by the liver in vivo. Pretreatment of the rats with chloroquine resulted in a decrease in proteolysis of the iodopeptide within the isolated vesicles.

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Selected References

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

  1. Anderson R. G., Brown M. S., Beisiegel U., Goldstein J. L. Surface distribution and recycling of the low density lipoprotein receptor as visualized with antireceptor antibodies. J Cell Biol. 1982 Jun;93(3):523–531. doi: 10.1083/jcb.93.3.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ascoli M., Puett D. Degradation of receptor-bound human choriogonadotropin by murine Leydig tumor cells. J Biol Chem. 1978 Jul 25;253(14):4892–4899. [PubMed] [Google Scholar]
  3. Brush J. S., Jering H. The importance of proteolysis as the initial step of insulin degradation in rat liver homogenates. Endocrinology. 1979 Jun;104(6):1639–1643. doi: 10.1210/endo-104-6-1639. [DOI] [PubMed] [Google Scholar]
  4. Burghen G. A., Kitabchi A. E., Brush J. S. Characterization of a rat liver protease with specificity for insulin. Endocrinology. 1972 Sep;91(3):633–642. doi: 10.1210/endo-91-3-633. [DOI] [PubMed] [Google Scholar]
  5. Campbell C. H., Rome L. H. Coated vesicles from rat liver and calf brain contain lysosomal enzymes bound to mannose 6-phosphate receptors. J Biol Chem. 1983 Nov 10;258(21):13347–13352. [PubMed] [Google Scholar]
  6. Draznin B., Solomons C. C., Toothaker D. R., Sussman K. E. Energy-dependent steps in insulin-hepatocyte interaction. Endocrinology. 1981 Jan;108(1):8–17. doi: 10.1210/endo-108-1-8. [DOI] [PubMed] [Google Scholar]
  7. Duckworth W. C., Runyan K. R., Wright R. K., Halban P. A., Solomon S. S. Insulin degradation by hepatocytes in primary culture. Endocrinology. 1981 Apr;108(4):1142–1147. doi: 10.1210/endo-108-4-1142. [DOI] [PubMed] [Google Scholar]
  8. Farquhar M. G. Multiple pathways of exocytosis, endocytosis, and membrane recycling: validation of a Golgi route. Fed Proc. 1983 May 15;42(8):2407–2413. [PubMed] [Google Scholar]
  9. Gliemann J., Sonne O. Binding and receptor-mediated degradation of insulin in adipocytes. J Biol Chem. 1978 Nov 10;253(21):7857–7863. [PubMed] [Google Scholar]
  10. Jones R. H., Sönksen P. H., Boroujerdi M. A., Carson E. R. Number and affinity of insulin receptors in intact human subjects. Diabetologia. 1984 Aug;27(2):207–211. doi: 10.1007/BF00273808. [DOI] [PubMed] [Google Scholar]
  11. Juul S. M., Jones R. H. Evidence for a direct effect of bacitracin on cell-mediated insulin degradation in isolated hepatocytes. Biochem J. 1982 Aug 15;206(2):295–299. doi: 10.1042/bj2060295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jørgensen K. H., Larsen U. D. Homogeneous mono-(125)i-insulins. Preparation and characterization of mono-(125)i-(tyr a14)-and mono-(125)i-(tyr a19)-insulin. Diabetologia. 1980;19(6):546–554. doi: 10.1007/BF00253183. [DOI] [PubMed] [Google Scholar]
  13. Khan M. N., Posner B. I., Khan R. J., Bergeron J. J. Internalization of insulin into rat liver Golgi elements. Evidence for vesicle heterogeneity and the path of intracellular processing. J Biol Chem. 1982 May 25;257(10):5969–5976. [PubMed] [Google Scholar]
  14. Leighton F., Poole B., Beaufay H., Baudhuin P., Coffey J. W., Fowler S., De Duve C. The large-scale separation of peroxisomes, mitochondria, and lysosomes from the livers of rats injected with triton WR-1339. Improved isolation procedures, automated analysis, biochemical and morphological properties of fractions. J Cell Biol. 1968 May;37(2):482–513. doi: 10.1083/jcb.37.2.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Misbin R. I., Almira E. C., Buynitzky S. J. Insulin metabolism in rat hepatocytes. Evidence for generation of an insulin fragment missing a portion of the B chain involved in receptor binding. J Biol Chem. 1983 Feb 25;258(4):2157–2162. [PubMed] [Google Scholar]
  16. Neal G. W., Solomon S. S., Shankar T. P., Duckworth W. C. Glucagon degradation by human mononuclear cells. Diabetologia. 1983 Nov;25(5):404–410. doi: 10.1007/BF00282519. [DOI] [PubMed] [Google Scholar]
  17. Palade G. E. Problems in intracellular membrane traffic. Ciba Found Symp. 1982;(92):1–14. doi: 10.1002/9780470720745.ch1. [DOI] [PubMed] [Google Scholar]
  18. Pease R. J., Sharp G. A., Smith G. D., Peters T. J. Isolation and partial characterization of the rat liver ligandosome fraction. Biochim Biophys Acta. 1984 Jul 11;774(1):56–66. doi: 10.1016/0005-2736(84)90274-8. [DOI] [PubMed] [Google Scholar]
  19. Pilch P. F., Shia M. A., Benson R. J., Fine R. E. Coated vesicles participate in the receptor-mediated endocytosis of insulin. J Cell Biol. 1983 Jan;96(1):133–138. doi: 10.1083/jcb.96.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Plas C., Desbuquois B. Receptor-mediated insulin degradation and insulin-stimulated glycogenesis in cultured foetal hepatocytes. Biochem J. 1982 Feb 15;202(2):333–341. doi: 10.1042/bj2020333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Poole G. P., O'Connor K. J., Lazarus N. R., Pogson C. I. 125I-labelled insulin degradation by isolated rat hepatocytes: the roles of glutathione-insulin transhydrogenase and insulin-specific protease. Diabetologia. 1982 Jul;23(1):49–53. doi: 10.1007/BF00257731. [DOI] [PubMed] [Google Scholar]
  22. Posner B. I., Bergeron J. J., Josefsberg Z., Khan M. N., Khan R. J., Patel B. A., Sikstrom R. A., Verma A. K. Polypeptide hormones: intracellular receptors and internalization. Recent Prog Horm Res. 1981;37:539–582. doi: 10.1016/b978-0-12-571137-1.50016-5. [DOI] [PubMed] [Google Scholar]
  23. Posner B. I., Patel B. A., Khan M. N., Bergeron J. J. Effect of chloroquine on the internalization of 125I-insulin into subcellular fractions of rat liver. Evidence for an effect of chloroquine on Golgi elements. J Biol Chem. 1982 May 25;257(10):5789–5799. [PubMed] [Google Scholar]
  24. Quinn P. S., Judah J. D. Calcium-dependent Golgi-vesicle fusion and cathepsin B in the conversion of proalbumin into albumin in rat liver. Biochem J. 1978 May 15;172(2):301–309. doi: 10.1042/bj1720301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Regoeczi E., Chindemi P. A., Debanne M. T., Charlwood P. A. Partial resialylation of human asialotransferrin type 3 in the rat. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2226–2230. doi: 10.1073/pnas.79.7.2226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schacterle G. R., Pollack R. L. A simplified method for the quantitative assay of small amounts of protein in biologic material. Anal Biochem. 1973 Feb;51(2):654–655. doi: 10.1016/0003-2697(73)90523-x. [DOI] [PubMed] [Google Scholar]
  27. Shah T., Heywood-Waddington D., Smith G. D., Peters T. J. Automated enzymic, protein and DNA microanalysis to tissue biopsy samples and subcellular fractions. Clin Chim Acta. 1984 Mar 27;138(1):125–132. doi: 10.1016/0009-8981(84)90361-9. [DOI] [PubMed] [Google Scholar]
  28. Smith G. D., Peters T. J. Analytical subcellular fractionation of rat liver with special reference to the localisation of putative plasma membrane marker enzymes. Eur J Biochem. 1980 Feb;104(1):305–311. doi: 10.1111/j.1432-1033.1980.tb04429.x. [DOI] [PubMed] [Google Scholar]
  29. Smith G. D., Peters T. J. The localization in rat liver of alkaline phosphodiesterase to a discrete organelle implicated in ligand internalization. Biochim Biophys Acta. 1982 May 5;716(1):24–30. doi: 10.1016/0304-4165(82)90198-2. [DOI] [PubMed] [Google Scholar]
  30. Terris S., Steiner D. F. Binding and degradation of 125I-insulin by rat hepatocytes. J Biol Chem. 1975 Nov 10;250(21):8389–8398. [PubMed] [Google Scholar]
  31. Terris S., Steiner D. F. Retention and degradation of 125I-insulin by perfused livers from diabetic rats. J Clin Invest. 1976 Apr;57(4):885–896. doi: 10.1172/JCI108365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tycko B., Maxfield F. R. Rapid acidification of endocytic vesicles containing alpha 2-macroglobulin. Cell. 1982 Mar;28(3):643–651. doi: 10.1016/0092-8674(82)90219-7. [DOI] [PubMed] [Google Scholar]
  33. Wall D. A., Wilson G., Hubbard A. L. The galactose-specific recognition system of mammalian liver: the route of ligand internalization in rat hepatocytes. Cell. 1980 Aug;21(1):79–93. doi: 10.1016/0092-8674(80)90116-6. [DOI] [PubMed] [Google Scholar]
  34. Welinder B. S., Linde S., Brush J. S. Separation of the monoiodinated isomers of insulin by high-performance liquid chromatography. J Chromatogr. 1983 Feb 25;257(1):162–165. doi: 10.1016/s0021-9673(01)88169-3. [DOI] [PubMed] [Google Scholar]
  35. Yokono K., Roth R. A., Baba S. Identification of insulin-degrading enzyme on the surface of cultured human lymphocytes, rat hepatoma cells, and primary cultures of rat hepatocytes. Endocrinology. 1982 Oct;111(4):1102–1108. doi: 10.1210/endo-111-4-1102. [DOI] [PubMed] [Google Scholar]

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