Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 May;81(10):3108–3112. doi: 10.1073/pnas.81.10.3108

Clathrin-coated vesicles from rat liver: enzymatic profile and characterization of ATP-dependent proton transport.

R W Van Dyke, C J Steer, B F Scharschmidt
PMCID: PMC345230  PMID: 6145154

Abstract

Clathrin-coated vesicles isolated from rat liver exhibited an enzymatic profile distinct from that of rat liver plasma membranes, lysosomes, microsomes, and mitochondria. The coated vesicles catalyzed ATP-dependent proton transport that acidified the vesicle interior, as measured by the fluorescence quenching of acridine orange. H+ transport by coated vesicles was not inhibited by vanadate (0.1 mM) or ouabain (2 mM) and differed from H+ transport by rat liver submitochondrial particles in its greater resistance to inhibition by oligomycin (10 pM to 10 microM) and N,N'-dicyclohexylcarbodiimide (DCCD) (0.1-100 microM) and its sensitivity to N-ethylmaleimide (0.1-2 mM). H+ transport was stimulated by valinomycin in the presence of K+, exhibited no specific cation requirement, but was dependent upon the presence of a permeant anion, with Cl- and Br- being the most effective of the anions studied. Finally, H+ transport was poorly supported by GTP, UTP, or ADP and exhibited no consistent relationship to the coated vesicle-associated ouabain-insensitive ATPase activity.

Full text

PDF
3108

Selected References

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

  1. Blitzer B. L., Boyer J. L. Cytochemical localization of Na+, K+-ATPase in the rat hepatocyte. J Clin Invest. 1978 Nov;62(5):1104–1108. doi: 10.1172/JCI109216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowman B. J., Mainzer S. E., Allen K. E., Slayman C. W. Effects of inhibitors on the plasma membrane and mitochondrial adenosine triphosphatases of Neurospora crassa. Biochim Biophys Acta. 1978 Sep 11;512(1):13–28. doi: 10.1016/0005-2736(78)90214-6. [DOI] [PubMed] [Google Scholar]
  3. Bowman B. J., Slayman C. W. Characterization of plasma membrane adenosine triphosphatase of Neurospora crassa. J Biol Chem. 1977 May 25;252(10):3357–3363. [PubMed] [Google Scholar]
  4. COOPERSTEIN S. J., LAZAROW A. A microspectrophotometric method for the determination of cytochrome oxidase. J Biol Chem. 1951 Apr;189(2):665–670. [PubMed] [Google Scholar]
  5. Dufour J. P., Boutry M., Goffeau A. Plasma membrane ATPase of yeast. Comparative inhibition studies of the purified and membrane-bound enzymes. J Biol Chem. 1980 Jun 25;255(12):5735–5741. [PubMed] [Google Scholar]
  6. 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]
  7. GIBBS G. E., REIMER K. QUANTITATIVE MICRODETERMINATION OF ENZYMES IN SWEAT GLAND. 3. SUCCINIC DEHYDROGENASE IN CYSTIC FIBROSIS. Proc Soc Exp Biol Med. 1965 Jun;119:470–473. doi: 10.3181/00379727-119-30213. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Gluck S., Cannon C., Al-Awqati Q. Exocytosis regulates urinary acidification in turtle bladder by rapid insertion of H+ pumps into the luminal membrane. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4327–4331. doi: 10.1073/pnas.79.14.4327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kopecky J., Guerrieri F., Papa S. Interaction of dicyclohexylcarbodiimide with the proton-conducting pathway of mitochondrial H+-ATPase. Eur J Biochem. 1983 Mar 1;131(1):17–24. doi: 10.1111/j.1432-1033.1983.tb07226.x. [DOI] [PubMed] [Google Scholar]
  12. LaRusso N. F., Fowler S. Coordinate secretion of acid hydrolases in rat bile. J Clin Invest. 1979 Oct;64(4):948–954. doi: 10.1172/JCI109561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Latham P. S., Kashgarian M. The ultrastructural localization of transport ATPase in the rat liver at non-bile canalicular plasma membranes. Gastroenterology. 1979 May;76(5 Pt 1):988–996. [PubMed] [Google Scholar]
  14. Lee H. C., Breitbart H., Berman M., Forte J. G. Potassium-stimulated ATPase activity and hydrogen transport in gastric microsomal vesicles. Biochim Biophys Acta. 1979 May 3;553(1):107–131. doi: 10.1016/0005-2736(79)90034-8. [DOI] [PubMed] [Google Scholar]
  15. Lee H. C., Forte J. G. A study of H+ transport in gastric microsomal vesicles using fluorescent probes. Biochim Biophys Acta. 1978 Apr 4;508(2):339–356. doi: 10.1016/0005-2736(78)90336-x. [DOI] [PubMed] [Google Scholar]
  16. Maloney P. C. Energy coupling to ATP synthesis by the proton-translocating ATPase. J Membr Biol. 1982;67(1):1–12. doi: 10.1007/BF01868643. [DOI] [PubMed] [Google Scholar]
  17. Marsh M., Bolzau E., Helenius A. Penetration of Semliki Forest virus from acidic prelysosomal vacuoles. Cell. 1983 Mar;32(3):931–940. doi: 10.1016/0092-8674(83)90078-8. [DOI] [PubMed] [Google Scholar]
  18. Maxfield F. R. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982 Nov;95(2 Pt 1):676–681. doi: 10.1083/jcb.95.2.676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mitchell P., Moyle J. Acid-base titration across the membrane system of rat-liver mitochondria. Catalysis by uncouplers. Biochem J. 1967 Aug;104(2):588–600. doi: 10.1042/bj1040588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nandi P. K., Irace G., Van Jaarsveld P. P., Lippoldt R. E., Edelhoch H. Instability of coated vesicles in concentrated sucrose solutions. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5881–5885. doi: 10.1073/pnas.79.19.5881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Renston R. H., Jones A. L., Christiansen W. D., Hradek G. T., Underdown B. J. Evidence for a vesicular transport mechanism in hepatocytes for biliary secretion of immunoglobulin A. Science. 1980 Jun 13;208(4449):1276–1278. doi: 10.1126/science.7375938. [DOI] [PubMed] [Google Scholar]
  23. Scharschmidt B. F., Keeffe E. B., Blankenship N. M., Ockner R. K. Validation of a recording spectrophotometric method for measurement of membrane-associated Mg- and NaK-ATPase activity. J Lab Clin Med. 1979 May;93(5):790–799. [PubMed] [Google Scholar]
  24. Scharschmidt B. F., Keeffe E. B. Isolation of a rat liver plasma membrane fraction of probable canalicular origin. Preparative technique, enzymatic profile, composition, and solute transport. Biochim Biophys Acta. 1981 Sep 7;646(3):369–381. doi: 10.1016/0005-2736(81)90305-9. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Steer C. J., Klausner R. D. Clathrin-coated pits and coated vesicles: functional and structural studies. Hepatology. 1983 May-Jun;3(3):437–454. doi: 10.1002/hep.1840030326. [DOI] [PubMed] [Google Scholar]
  27. Steer C. J., Wall D. A., Ashwell G. Evidence for the presence of the asialoglycoprotein receptor in coated vesicles isolated from rat liver. Hepatology. 1983 Sep-Oct;3(5):667–672. doi: 10.1002/hep.1840030507. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Straubinger R. M., Hong K., Friend D. S., Papahadjopoulos D. Endocytosis of liposomes and intracellular fate of encapsulated molecules: encounter with a low pH compartment after internalization in coated vesicles. Cell. 1983 Apr;32(4):1069–1079. doi: 10.1016/0092-8674(83)90291-x. [DOI] [PubMed] [Google Scholar]
  30. Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Warnock D. G., Reenstra W. W., Yee V. J. Na+/H+ antiporter of brush border vesicles: studies with acridine orange uptake. Am J Physiol. 1982 Jun;242(6):F733–F739. doi: 10.1152/ajprenal.1982.242.6.F733. [DOI] [PubMed] [Google Scholar]
  33. Willsky G. R. Characterization of the plasma membrane Mg2+-ATPase from the yeast, Saccharomyces cerevisiae. J Biol Chem. 1979 May 10;254(9):3326–3332. [PubMed] [Google Scholar]
  34. Xie X. S., Stone D. K., Racker E. Determinants of clathrin-coated vesicle acidification. J Biol Chem. 1983 Dec 25;258(24):14834–14838. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES