The distribution of 215-kilodalton mannose 6-phosphate receptors within cis (heavy) and trans (light) Golgi subfractions varies in different cell types (original) (raw)

Abstract

The distribution of mannose 6-phosphate (Man-6-P) receptors for lysosomal enzymes was investigated in Golgi subfractions prepared from three different cultured cell lines. Total microsomal fractions from clone 9 hepatocytes, normal rat kidney, or Chinese hamster ovary cells were subfractionated by flotation in sucrose density gradients, which resolves Golgi membranes into heavy (cis), intermediate, and light (trans) subfractions. The distribution of Man-6-P receptors within the subfractions was assessed by quantitative immunoprecipitation, and the results were compared to those obtained by immunoperoxidase localization of the receptors in Golgi cisternae of intact cells. In all cases, the results obtained by Golgi subfractionation and by immunoelectron microscopy were in agreement. In clone 9 cells, Man-6-P receptors were enriched in heavy (cis) Golgi subfractions, whose peak density (rho = 1.17) was greater than those containing either galactosyltransferase activity, a trans Golgi marker, or alpha-mannosidase II, a middle Golgi marker. By immunoelectron microscopy, the receptors were localized to a single cis Golgi cisterna. In Chinese hamster ovary cells, Man-6-P receptors were concentrated in Golgi membranes of low density (1.12 g/ml) overlapping the peak of galactosyltransferase activity. By the immunoperoxidase technique, the receptors were usually localized to a single trans Golgi cisterna. In normal rat kidney cells, Man-6-P receptors were found to be broadly distributed across Golgi membranes (rho = 1.12-1.17), and by immunoperoxidase localization they were found to be broadly distributed across the stacked Golgi cisternae. It is concluded that the distribution of Man-6-P receptors within the Golgi complex varies from one cell type to another. These differences in receptor distribution may reflect variations in lysosomal enzyme trafficking among different cell types.

9001

Images in this article

Selected References

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

  1. Balch W. E., Rothman J. E. Characterization of protein transport between successive compartments of the Golgi apparatus: asymmetric properties of donor and acceptor activities in a cell-free system. Arch Biochem Biophys. 1985 Jul;240(1):413–425. doi: 10.1016/0003-9861(85)90046-3. [DOI] [PubMed] [Google Scholar]
  2. Brew K., Shaper J. H., Olsen K. W., Trayer I. P., Hill R. L. Cross-linking of the components of lactose synthetase with dimethylpimelimidate. J Biol Chem. 1975 Feb 25;250(4):1434–1444. [PubMed] [Google Scholar]
  3. Brown W. J., Constantinescu E., Farquhar M. G. Redistribution of mannose-6-phosphate receptors induced by tunicamycin and chloroquine. J Cell Biol. 1984 Jul;99(1 Pt 1):320–326. doi: 10.1083/jcb.99.1.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown W. J., Farquhar M. G. Accumulation of coated vesicles bearing mannose 6-phosphate receptors for lysosomal enzymes in the Golgi region of I-cell fibroblasts. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5135–5139. doi: 10.1073/pnas.81.16.5135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown W. J., Farquhar M. G. The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae. Cell. 1984 Feb;36(2):295–307. doi: 10.1016/0092-8674(84)90223-x. [DOI] [PubMed] [Google Scholar]
  6. Brown W. J., Goodhouse J., Farquhar M. G. Mannose-6-phosphate receptors for lysosomal enzymes cycle between the Golgi complex and endosomes. J Cell Biol. 1986 Oct;103(4):1235–1247. doi: 10.1083/jcb.103.4.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Chamberlain J. P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
  9. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dunphy W. G., Rothman J. E. Compartmentation of asparagine-linked oligosaccharide processing in the Golgi apparatus. J Cell Biol. 1983 Jul;97(1):270–275. doi: 10.1083/jcb.97.1.270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farquhar M. G., Palade G. E. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. doi: 10.1083/jcb.91.3.77s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Farquhar M. G. Progress in unraveling pathways of Golgi traffic. Annu Rev Cell Biol. 1985;1:447–488. doi: 10.1146/annurev.cb.01.110185.002311. [DOI] [PubMed] [Google Scholar]
  13. Fedde K. N., Sly W. S. Ricin-binding properties of acid hydrolases from isolated lysosomes implies prior processing by terminal transferases of the trans-Golgi apparatus. Biochem Biophys Res Commun. 1985 Dec 17;133(2):614–620. doi: 10.1016/0006-291x(85)90949-0. [DOI] [PubMed] [Google Scholar]
  14. Geuze H. J., Slot J. W., Strous G. J., Hasilik A., Von Figura K. Ultrastructural localization of the mannose 6-phosphate receptor in rat liver. J Cell Biol. 1984 Jun;98(6):2047–2054. doi: 10.1083/jcb.98.6.2047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Geuze H. J., Slot J. W., Strous G. J., Hasilik A., von Figura K. Possible pathways for lysosomal enzyme delivery. J Cell Biol. 1985 Dec;101(6):2253–2262. doi: 10.1083/jcb.101.6.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goldberg D. E., Kornfeld S. Evidence for extensive subcellular organization of asparagine-linked oligosaccharide processing and lysosomal enzyme phosphorylation. J Biol Chem. 1983 Mar 10;258(5):3159–3165. [PubMed] [Google Scholar]
  17. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  18. Hoflack B., Kornfeld S. Lysosomal enzyme binding to mouse P388D1 macrophage membranes lacking the 215-kDa mannose 6-phosphate receptor: evidence for the existence of a second mannose 6-phosphate receptor. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4428–4432. doi: 10.1073/pnas.82.13.4428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoflack B., Kornfeld S. Purification and characterization of a cation-dependent mannose 6-phosphate receptor from murine P388D1 macrophages and bovine liver. J Biol Chem. 1985 Oct 5;260(22):12008–12014. [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. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Miller A. L., Kress B. C., Stein R., Kinnon C., Kern H., Schneider J. A., Harms E. Properties of N-acetyl-beta-D-hexosaminidase from isolated normal and I-cell lysosomes. J Biol Chem. 1981 Sep 10;256(17):9352–9362. [PubMed] [Google Scholar]
  23. Minnifield N., Creek K. E., Navas P., Morré D. J. Involvement of cis and trans Golgi apparatus elements in the intracellular sorting and targeting of acid hydrolases to lysosomes. Eur J Cell Biol. 1986 Oct;42(1):92–100. [PubMed] [Google Scholar]
  24. Nakao Y., Kozutsumi Y., Kawasaki T., Yamashina I., Van Halbeek H., Vliegenthart J. F. Oligosaccharides on cathepsin D from porcine spleen. Arch Biochem Biophys. 1984 Feb 15;229(1):43–54. doi: 10.1016/0003-9861(84)90128-0. [DOI] [PubMed] [Google Scholar]
  25. Rosenfeld M. G., Kreibich G., Popov D., Kato K., Sabatini D. D. Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution. J Cell Biol. 1982 Apr;93(1):135–143. doi: 10.1083/jcb.93.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roth J., Berger E. G. Immunocytochemical localization of galactosyltransferase in HeLa cells: codistribution with thiamine pyrophosphatase in trans-Golgi cisternae. J Cell Biol. 1982 Apr;93(1):223–229. doi: 10.1083/jcb.93.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Roth J., Taatjes D. J., Lucocq J. M., Weinstein J., Paulson J. C. Demonstration of an extensive trans-tubular network continuous with the Golgi apparatus stack that may function in glycosylation. Cell. 1985 Nov;43(1):287–295. doi: 10.1016/0092-8674(85)90034-0. [DOI] [PubMed] [Google Scholar]
  28. Rothman J. E., Fries E. Transport of newly synthesized vesicular stomatitis viral glycoprotein to purified Golgi membranes. J Cell Biol. 1981 Apr;89(1):162–168. doi: 10.1083/jcb.89.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sahagian G. G., Distler J., Jourdian G. W. Characterization of a membrane-associated receptor from bovine liver that binds phosphomannosyl residues of bovine testicular beta-galactosidase. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4289–4293. doi: 10.1073/pnas.78.7.4289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sahagian G. G. The mannose 6-phosphate receptor: function, biosynthesis and translocation. Biol Cell. 1984;51(2):207–214. doi: 10.1111/j.1768-322x.1984.tb00300.x. [DOI] [PubMed] [Google Scholar]
  31. Snider M. D., Rogers O. C. Membrane traffic in animal cells: cellular glycoproteins return to the site of Golgi mannosidase I. J Cell Biol. 1986 Jul;103(1):265–275. doi: 10.1083/jcb.103.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Takahashi T., Schmidt P. G., Tang J. Oligosaccharide units of lysosomal cathepsin D from porcine spleen. Amino acid sequence and carbohydrate structure of the glycopeptides. J Biol Chem. 1983 Mar 10;258(5):2819–2830. [PubMed] [Google Scholar]
  33. Tulsiani D. R., Hubbard S. C., Robbins P. W., Touster O. alpha-D-Mannosidases of rat liver Golgi membranes. Mannosidase II is the GlcNAcMAN5-cleaving enzyme in glycoprotein biosynthesis and mannosidases Ia and IB are the enzymes converting Man9 precursors to Man5 intermediates. J Biol Chem. 1982 Apr 10;257(7):3660–3668. [PubMed] [Google Scholar]
  34. Willingham M. C., Pastan I. H., Sahagian G. G. Ultrastructural immunocytochemical localization of the phosphomannosyl receptor in Chinese hamster ovary (CHO) cells. J Histochem Cytochem. 1983 Jan;31(1):1–11. doi: 10.1177/31.1.6300218. [DOI] [PubMed] [Google Scholar]
  35. Woods J. W., Doriaux M., Farquhar M. G. Transferrin receptors recycle to cis and middle as well as trans Golgi cisternae in Ig-secreting myeloma cells. J Cell Biol. 1986 Jul;103(1):277–286. doi: 10.1083/jcb.103.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. von Figura K., Hasilik A. Lysosomal enzymes and their receptors. Annu Rev Biochem. 1986;55:167–193. doi: 10.1146/annurev.bi.55.070186.001123. [DOI] [PubMed] [Google Scholar]