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. 1985 Jan 1;225(1):51–58. doi: 10.1042/bj2250051

Hepatic endosome fractions contain an ATP-driven proton pump.

T Saermark, N Flint, W H Evans
PMCID: PMC1144552  PMID: 2983664

Abstract

Endosome fractions were isolated from rat liver homogenates on the basis of the subcellular distribution of circulating ligands, e.g. 125I-asialotransferrin internalized by hepatocytes by a receptor-mediated process. The distribution of endocytosed 125I-asialotransferrin 1-2 min and 15 min after uptake by liver and a monensin-activated Mg2+-dependent ATPase activity coincided on linear gradients of sucrose and Nycodenz. The monensin-activated Mg2+-ATPase was enriched relative to the liver homogenates up to 60-fold in specific activity in the endosome fractions. Contamination of the endosome fractions by lysosomes, endoplasmic reticulum, mitochondria, plasma membranes and Golgi-apparatus components was low. By use of 9-aminoacridine, a probe for pH gradients, the endosome vesicles were shown to acidify on addition of ATP. Acidification was reversed by addition of monensin. The results indicate that endosome fractions contain an ATP-driven proton pump. The ionophore-activated Mg2+-ATPase in combination with the presence of undegraded ligands in the endosome fractions emerge as linked markers for this new subcellular organelle.

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Selected References

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

  1. Bergeron J. J. Golgi fractions from livers of control and ethanol-intoxicated rats. Enzymic and morphologic properties following rapid isolation. Biochim Biophys Acta. 1979 Aug 23;555(3):493–503. doi: 10.1016/0005-2736(79)90402-4. [DOI] [PubMed] [Google Scholar]
  2. Bergeron J. J., Rachubinski R. A., Sikstrom R. A., Posner B. I., Paiement J. Galactose transfer to endogenous acceptors within Golgi fractions of rat liver. J Cell Biol. 1982 Jan;92(1):139–146. doi: 10.1083/jcb.92.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blitz A. L., Fine R. E., Toselli P. A. Evidence that coated vesicles isolated from brain are calcium-sequestering organelles resembling sarcoplasmic reticulum. J Cell Biol. 1977 Oct;75(1):135–147. doi: 10.1083/jcb.75.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown M. S., Anderson R. G., Goldstein J. L. Recycling receptors: the round-trip itinerary of migrant membrane proteins. Cell. 1983 Mar;32(3):663–667. doi: 10.1016/0092-8674(83)90052-1. [DOI] [PubMed] [Google Scholar]
  5. Cidon S., Nelson N. A novel ATPase in the chromaffin granule membrane. J Biol Chem. 1983 Mar 10;258(5):2892–2898. [PubMed] [Google Scholar]
  6. Debanne M. T., Evans W. H., Flint N., Regoeczi E. Receptor-rich intracellular membrane vesicles transporting asialotransferrin and insulin in liver. Nature. 1982 Jul 22;298(5872):398–400. doi: 10.1038/298398a0. [DOI] [PubMed] [Google Scholar]
  7. Desbuquois B., Willeput J., Huet de Froberville A. Receptor-mediated internalisation of insulin in intact rat liver. A biochemical study. FEBS Lett. 1979 Oct 15;106(2):338–344. doi: 10.1016/0014-5793(79)80528-1. [DOI] [PubMed] [Google Scholar]
  8. ENGSTROM L. Studies on calf-intestinal alkaline phosphatase. I. Chromatographic purification, microheterogeneity and some other properties of the purified enzyme. Biochim Biophys Acta. 1961 Sep 2;52:36–48. doi: 10.1016/0006-3002(61)90901-5. [DOI] [PubMed] [Google Scholar]
  9. Evans W. H. Preparation of low-density "endosome" and "endosome"-depleted Golgi fractions from rat liver. Methods Enzymol. 1985;109:246–257. doi: 10.1016/0076-6879(85)09090-5. [DOI] [PubMed] [Google Scholar]
  10. Famulski K., Carafoli E. Ca2+ transporting activity of membrane fractions isolated from the post-mitochondrial supernatant of rat liver. Cell Calcium. 1982 Aug;3(3):263–281. doi: 10.1016/0143-4160(82)90005-7. [DOI] [PubMed] [Google Scholar]
  11. Forgac M., Cantley L., Wiedenmann B., Altstiel L., Branton D. Clathrin-coated vesicles contain an ATP-dependent proton pump. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1300–1303. doi: 10.1073/pnas.80.5.1300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Galloway C. J., Dean G. E., Marsh M., Rudnick G., Mellman I. Acidification of macrophage and fibroblast endocytic vesicles in vitro. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3334–3338. doi: 10.1073/pnas.80.11.3334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Geisow M. J., Evans W. H. pH in the endosome. Measurements during pinocytosis and receptor-mediated endocytosis. Exp Cell Res. 1984 Jan;150(1):36–46. doi: 10.1016/0014-4827(84)90699-2. [DOI] [PubMed] [Google Scholar]
  14. Geuze H. J., Slot J. W., Strous G. J., Lodish H. F., Schwartz A. L. Intracellular site of asialoglycoprotein receptor-ligand uncoupling: double-label immunoelectron microscopy during receptor-mediated endocytosis. Cell. 1983 Jan;32(1):277–287. doi: 10.1016/0092-8674(83)90518-4. [DOI] [PubMed] [Google Scholar]
  15. Glickman J., Croen K., Kelly S., Al-Awqati Q. Golgi membranes contain an electrogenic H+ pump in parallel to a chloride conductance. J Cell Biol. 1983 Oct;97(4):1303–1308. doi: 10.1083/jcb.97.4.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Harford J., Bridges K., Ashwell G., Klausner R. D. Intracellular dissociation of receptor-bound asialoglycoproteins in cultured hepatocytes. A pH-mediated nonlysosomal event. J Biol Chem. 1983 Mar 10;258(5):3191–3197. [PubMed] [Google Scholar]
  17. Harikumar P., Reeves J. P. The lysosomal proton pump is electrogenic. J Biol Chem. 1983 Sep 10;258(17):10403–10410. [PubMed] [Google Scholar]
  18. Hopkins C. R. Intracellular routing of transferrin and transferrin receptors in epidermoid carcinoma A431 cells. Cell. 1983 Nov;35(1):321–330. doi: 10.1016/0092-8674(83)90235-0. [DOI] [PubMed] [Google Scholar]
  19. Kinsky S. C. Antibiotic interaction with model membranes. Annu Rev Pharmacol. 1970;10:119–142. doi: 10.1146/annurev.pa.10.040170.001003. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Luzio J. P., Stanley K. K. The isolation of endosome-derived vesicles from rat hepatocytes. Biochem J. 1983 Oct 15;216(1):27–36. doi: 10.1042/bj2160027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Merion M., Schlesinger P., Brooks R. M., Moehring J. M., Moehring T. J., Sly W. S. Defective acidification of endosomes in Chinese hamster ovary cell mutants "cross-resistant" to toxins and viruses. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5315–5319. doi: 10.1073/pnas.80.17.5315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mrsny R. J., Meizel S. Potassium ion influx and Na+,K+-ATPase activity are required for the hamster sperm acrosome reaction. J Cell Biol. 1981 Oct;91(1):77–82. [PMC free article] [PubMed] [Google Scholar]
  24. Ohkuma S., Moriyama Y., Takano T. Identification and characterization of a proton pump on lysosomes by fluorescein-isothiocyanate-dextran fluorescence. Proc Natl Acad Sci U S A. 1982 May;79(9):2758–2762. doi: 10.1073/pnas.79.9.2758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pastan I. H., Willingham M. C. Journey to the center of the cell: role of the receptosome. Science. 1981 Oct 30;214(4520):504–509. doi: 10.1126/science.6170111. [DOI] [PubMed] [Google Scholar]
  26. Pearse B. M., Bretscher M. S. Membrane recycling by coated vesicles. Annu Rev Biochem. 1981;50:85–101. doi: 10.1146/annurev.bi.50.070181.000505. [DOI] [PubMed] [Google Scholar]
  27. Pricer W. E., Jr, Ashwell G. Subcellular distribution of a mammalian hepatic binding protein specific for asialoglycoproteins. J Biol Chem. 1976 Dec 10;251(23):7539–7544. [PubMed] [Google Scholar]
  28. 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]
  29. Regoeczi E., Chindemi P. A., Debanne M. T., Hatton M. W. Dual nature of the hepatic lectin pathway for human asialotransferrin type 3 in the rat. J Biol Chem. 1982 May 25;257(10):5431–5436. [PubMed] [Google Scholar]
  30. Saermark T., Jones P. M., Robinson I. C. Membrane retrieval in the guinea-pig neurohypophysis. Isolation and characterization of secretory vesicles and coated microvesicles after radiolabel incorporation in vivo. Biochem J. 1984 Mar 1;218(2):591–599. doi: 10.1042/bj2180591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saermark T., Thorn N. A. Ca2+-Mg2+-ATPase activity in brain coated microvesicles purified on immunosorbents. Cell Calcium. 1982 Dec;3(6):561–581. doi: 10.1016/0143-4160(82)90045-8. [DOI] [PubMed] [Google Scholar]
  32. Saermark T., Vilhardt H. Isolation and partial characterization of magnesium ion- and calcium ion-dependent adenosine triphosphatase activity from bovine brain microsomal fraction. Biochem J. 1979 Aug 1;181(2):321–330. doi: 10.1042/bj1810321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schneider D. L. ATP-dependent acidification of intact and disrupted lysosomes. Evidence for an ATP-driven proton pump. J Biol Chem. 1981 Apr 25;256(8):3858–3864. [PubMed] [Google Scholar]
  34. Schuldiner S., Rottenberg H., Avron M. Determination of pH in chloroplasts. 2. Fluorescent amines as a probe for the determination of pH in chloroplasts. Eur J Biochem. 1972 Jan 31;25(1):64–70. doi: 10.1111/j.1432-1033.1972.tb01667.x. [DOI] [PubMed] [Google Scholar]
  35. Stone D. K., Xie X. S., Racker E. An ATP-driven proton pump in clathrin-coated vesicles. J Biol Chem. 1983 Apr 10;258(7):4059–4062. [PubMed] [Google Scholar]
  36. 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]
  37. Vischer P., Reutter W. Specific alterations of fucoprotein biosynthesis in the plasma membrane of Morris hepatoma 7777. Eur J Biochem. 1978 Mar 15;84(2):363–368. doi: 10.1111/j.1432-1033.1978.tb12176.x. [DOI] [PubMed] [Google Scholar]
  38. Wattiaux R., Wattiaux-De Coninck S., Ronveaux-dupal M. F., Dubois F. Isolation of rat liver lysosomes by isopycnic centrifugation in a metrizamide gradient. J Cell Biol. 1978 Aug;78(2):349–368. doi: 10.1083/jcb.78.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wisher M. H., Evans W. H. Functional polarity of the rat hepatocyte surface membrane. Isolation and characterization of plasma-membrane subfractions from the blood-sinusoidal, bile-Canalicular and contiguous surfaces of the hepatocyte. Biochem J. 1975 Feb;146(2):375–388. doi: 10.1042/bj1460375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yamashiro D. J., Fluss S. R., Maxfield F. R. Acidification of endocytic vesicles by an ATP-dependent proton pump. J Cell Biol. 1983 Sep;97(3):929–934. doi: 10.1083/jcb.97.3.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zhang F., Schneider D. L. The bioenergetics of Golgi apparatus function: evidence for an ATP-dependent proton pump. Biochem Biophys Res Commun. 1983 Jul 29;114(2):620–625. doi: 10.1016/0006-291x(83)90825-2. [DOI] [PubMed] [Google Scholar]
  42. van Deurs B., Christensen E. I. Endocytosis in kidney proximal tubule cells and cultured fibroblasts: a review of the structural aspects of membrane recycling between the plasma membrane and endocytic vacuoles. Eur J Cell Biol. 1984 Jan;33(1):163–173. [PubMed] [Google Scholar]
  43. van Renswoude J., Bridges K. R., Harford J. B., Klausner R. D. Receptor-mediated endocytosis of transferrin and the uptake of fe in K562 cells: identification of a nonlysosomal acidic compartment. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6186–6190. doi: 10.1073/pnas.79.20.6186. [DOI] [PMC free article] [PubMed] [Google Scholar]

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