• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. Dec 2, 1991; 115(6): 1505–1519.
PMCID: PMC2289203

Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network

Abstract

Regulated secretory proteins are thought to be sorted in the trans- Golgi network (TGN) via selective aggregation. The factors responsible for this aggregation are unknown. We show here that two widespread regulated secretory proteins, chromogranin B and secretogranin II (granins), remain in an aggregated state when TGN vesicles from neuroendocrine cells (PC12) are permeabilized at pH 6.4 in 1-10 mM calcium, conditions believed to exist in this compartment. Permeabilization of immature secretory granules under these conditions allowed the recovery of electron dense cores. The granin aggregates in the TGN largely excluded glycosaminoglycan chains which served as constitutively secreted bulk flow markers. The low pH, high calcium milieu was sufficient to induce granin aggregation in the RER. In the TGN of pituitary GH4C1 cells, the proportion of granins conserved as aggregates was higher upon hormonal treatment known to increase secretory granule formation. Our data suggest that a decrease in pH and an increase in calcium are sufficient to trigger the selective aggregation of the granins in the TGN, segregating them from constitutive secretory proteins.

Full Text

The Full Text of this article is available as a PDF (2.6M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson RG, Orci L. A view of acidic intracellular compartments. J Cell Biol. 1988 Mar;106(3):539–543. [PMC free article] [PubMed]
  • Baeuerle PA, Huttner WB. Tyrosine sulfation is a trans-Golgi-specific protein modification. J Cell Biol. 1987 Dec;105(6 Pt 1):2655–2664. [PMC free article] [PubMed]
  • Bassetti M, Huttner WB, Zanini A, Rosa P. Co-localization of secretogranins/chromogranins with thyrotropin and luteinizing hormone in secretory granules of cow anterior pituitary. J Histochem Cytochem. 1990 Sep;38(9):1353–1363. [PubMed]
  • Bendayan M. Concentration of amylase along its secretory pathway in the pancreatic acinar cell as revealed by high resolution immunocytochemistry. Histochem J. 1984 Jan;16(1):85–108. [PubMed]
  • Bendayan M, Roth J, Perrelet A, Orci L. Quantitative immunocytochemical localization of pancreatic secretory proteins in subcellular compartments of the rat acinar cell. J Histochem Cytochem. 1980 Feb;28(2):149–160. [PubMed]
  • Benedum UM, Baeuerle PA, Konecki DS, Frank R, Powell J, Mallet J, Huttner WB. The primary structure of bovine chromogranin A: a representative of a class of acidic secretory proteins common to a variety of peptidergic cells. EMBO J. 1986 Jul;5(7):1495–1502. [PMC free article] [PubMed]
  • Benedum UM, Lamouroux A, Konecki DS, Rosa P, Hille A, Baeuerle PA, Frank R, Lottspeich F, Mallet J, Huttner WB. The primary structure of human secretogranin I (chromogranin B): comparison with chromogranin A reveals homologous terminal domains and a large intervening variable region. EMBO J. 1987 May;6(5):1203–1211. [PMC free article] [PubMed]
  • Bulenda D, Gratzl M. Matrix free Ca2+ in isolated chromaffin vesicles. Biochemistry. 1985 Dec 17;24(26):7760–7765. [PubMed]
  • Burgess TL, Kelly RB. Sorting and secretion of adrenocorticotropin in a pituitary tumor cell line after perturbation of the level of a secretory granule-specific proteoglycan. J Cell Biol. 1984 Dec;99(6):2223–2230. [PMC free article] [PubMed]
  • Burgess TL, Kelly RB. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. [PubMed]
  • Cozzi MG, Zanini A. Secretogranin II is a Ca2+-binding protein. Cell Biol Int Rep. 1988 Jun;12(6):493–493. [PubMed]
  • Farquhar MG, Palade GE. The Golgi apparatus (complex)-(1954-1981)-from artifact to center stage. J Cell Biol. 1981 Dec;91(3 Pt 2):77s–103s. [PMC free article] [PubMed]
  • Fisher JM, Sossin W, Newcomb R, Scheller RH. Multiple neuropeptides derived from a common precursor are differentially packaged and transported. Cell. 1988 Sep 9;54(6):813–822. [PubMed]
  • Gerdes HH, Rosa P, Phillips E, Baeuerle PA, Frank R, Argos P, Huttner WB. The primary structure of human secretogranin II, a widespread tyrosine-sulfated secretory granule protein that exhibits low pH- and calcium-induced aggregation. J Biol Chem. 1989 Jul 15;264(20):12009–12015. [PubMed]
  • Giannattasio G, Zanini A, Meldolesi J. Molecular organization of rat prolactin granules. I. In vitro stability of intact and "membraneless" granules. J Cell Biol. 1975 Jan;64(1):246–251. [PMC free article] [PubMed]
  • Gorr SU, Shioi J, Cohn DV. Interaction of calcium with porcine adrenal chromogranin A (secretory protein-I) and chromogranin B (secretogranin I). Am J Physiol. 1989 Aug;257(2 Pt 1):E247–E254. [PubMed]
  • 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. [PubMed]
  • Hashimoto S, Fumagalli G, Zanini A, Meldolesi J. Sorting of three secretory proteins to distinct secretory granules in acidophilic cells of cow anterior pituitary. J Cell Biol. 1987 Oct;105(4):1579–1586. [PMC free article] [PubMed]
  • Huttner WB, Tooze SA. Biosynthetic protein transport in the secretory pathway. Curr Opin Cell Biol. 1989 Aug;1(4):648–654. [PubMed]
  • Huttner WB, Gerdes HH, Rosa P. The granin (chromogranin/secretogranin) family. Trends Biochem Sci. 1991 Jan;16(1):27–30. [PubMed]
  • Iacangelo A, Affolter HU, Eiden LE, Herbert E, Grimes M. Bovine chromogranin A sequence and distribution of its messenger RNA in endocrine tissues. Nature. 1986 Sep 4;323(6083):82–86. [PubMed]
  • Johnson RG., Jr Proton pumps and chemiosmotic coupling as a generalized mechanism for neurotransmitter and hormone transport. Ann N Y Acad Sci. 1987;493:162–177. [PubMed]
  • Kelly RB. Pathways of protein secretion in eukaryotes. Science. 1985 Oct 4;230(4721):25–32. [PubMed]
  • Kiino DR, Dannies PS. Hormonal regulation of prolactin storage in a clonal strain of rat pituitary tumor cells. Yale J Biol Med. 1982 Sep-Dec;55(5-6):409–420. [PMC free article] [PubMed]
  • Kimura JH, Lohmander LS, Hascall VC. Studies on the biosynthesis of cartilage proteoglycan in a model system of cultured chondrocytes from the Swarm rat chondrosarcoma. J Cell Biochem. 1984;26(4):261–278. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lohmander LS, Hascall VC, Caplan AI. Effects of 4-methyl umbelliferyl-beta-D-xylopyranoside on chondrogenesis and proteoglycan synthesis in chick limb bud mesenchymal cell cultures. J Biol Chem. 1979 Oct 25;254(20):10551–10561. [PubMed]
  • Macer DR, Koch GL. Identification of a set of calcium-binding proteins in reticuloplasm, the luminal content of the endoplasmic reticulum. J Cell Sci. 1988 Sep;91(Pt 1):61–70. [PubMed]
  • Mata M, Fink DJ. Ca++-ATPase in the central nervous system: an EM cytochemical study. J Histochem Cytochem. 1989 Jul;37(7):971–980. [PubMed]
  • Mata M, Staple J, Fink DJ. Ultrastructural distribution of Ca++ within neurons. An oxalate pyroantimonate study. Histochemistry. 1987;87(4):339–349. [PubMed]
  • Meldolesi J, Madeddu L, Pozzan T. Intracellular Ca2+ storage organelles in non-muscle cells: heterogeneity and functional assignment. Biochim Biophys Acta. 1990 Nov 12;1055(2):130–140. [PubMed]
  • Orci L, Ravazzola M, Amherdt M, Perrelet A, Powell SK, Quinn DL, Moore HP. The trans-most cisternae of the Golgi complex: a compartment for sorting of secretory and plasma membrane proteins. Cell. 1987 Dec 24;51(6):1039–1051. [PubMed]
  • Pfeffer SR, Rothman JE. Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi. Annu Rev Biochem. 1987;56:829–852. [PubMed]
  • Posthuma G, Slot JW, Veenendaal T, Geuze HJ. Immunogold determination of amylase concentrations in pancreatic subcellular compartments. Eur J Cell Biol. 1988 Jun;46(2):327–335. [PubMed]
  • Ravazzola M. Intracellular localization of calcium in the chromaffin cells of the rat adrenal medulla. Endocrinology. 1976 Apr;98(4):950–953. [PubMed]
  • Reiffen FU, Gratzl M. Chromogranins, widespread in endocrine and nervous tissue, bind Ca2+. FEBS Lett. 1986 Jan 20;195(1-2):327–330. [PubMed]
  • Roos N. A possible site of calcium regulation in rat exocrine pancreas cells: an X-ray microanalytical study. Scanning Microsc. 1988 Mar;2(1):323–329. [PubMed]
  • Rosa P, Hille A, Lee RW, Zanini A, De Camilli P, Huttner WB. Secretogranins I and II: two tyrosine-sulfated secretory proteins common to a variety of cells secreting peptides by the regulated pathway. J Cell Biol. 1985 Nov;101(5 Pt 1):1999–2011. [PMC free article] [PubMed]
  • Rosa P, Weiss U, Pepperkok R, Ansorge W, Niehrs C, Stelzer EH, Huttner WB. An antibody against secretogranin I (chromogranin B) is packaged into secretory granules. J Cell Biol. 1989 Jul;109(1):17–34. [PMC free article] [PubMed]
  • Salpeter MM, Farquhar MG. High resolution analysis of the secretory pathway in mammotrophs of the rat anterior pituitary. J Cell Biol. 1981 Oct;91(1):240–246. [PMC free article] [PubMed]
  • Sambrook JF. The involvement of calcium in transport of secretory proteins from the endoplasmic reticulum. Cell. 1990 Apr 20;61(2):197–199. [PubMed]
  • Scammell JG, Burrage TG, Dannies PS. Hormonal induction of secretory granules in a pituitary tumor cell line. Endocrinology. 1986 Oct;119(4):1543–1548. [PubMed]
  • Scammell JG, Rosa P, Hille A, Huttner WB. Regulation of chromogranin B/secretogranin I and secretogranin II storage in GH4C1 cells. J Histochem Cytochem. 1990 Jul;38(7):949–956. [PubMed]
  • Sossin WS, Fisher JM, Scheller RH. Sorting within the regulated secretory pathway occurs in the trans-Golgi network. J Cell Biol. 1990 Jan;110(1):1–12. [PMC free article] [PubMed]
  • Stoeckel ME, Hindelang-Gertner C, Dellmann H-D, Porte A, Stutinsky F. Subcellular localization of calcium in the mouse hypophysis. I. Calcium distribution in the adeno- and neurohypophysis under normal conditions. Cell Tissue Res. 1975;157(3):307–322. [PubMed]
  • Tashjian AH., Jr Clonal strains of hormone-producing pituitary cells. Methods Enzymol. 1979;58:527–535. [PubMed]
  • Tooze SA, Huttner WB. Cell-free protein sorting to the regulated and constitutive secretory pathways. Cell. 1990 Mar 9;60(5):837–847. [PubMed]
  • Tooze J, Tooze SA, Fuller SD. Sorting of progeny coronavirus from condensed secretory proteins at the exit from the trans-Golgi network of AtT20 cells. J Cell Biol. 1987 Sep;105(3):1215–1226. [PMC free article] [PubMed]
  • Tooze J, Kern HF, Fuller SD, Howell KE. Condensation-sorting events in the rough endoplasmic reticulum of exocrine pancreatic cells. J Cell Biol. 1989 Jul;109(1):35–50. [PMC free article] [PubMed]
  • Tooze SA, Flatmark T, Tooze J, Huttner WB. Characterization of the immature secretory granule, an intermediate in granule biogenesis. J Cell Biol. 1991 Dec;115(6):1491–1503. [PMC free article] [PubMed]
  • Tougard C, Nasciutti LE, Picart R, Tixier-Vidal A, Huttner WB. Subcellular distribution of secretogranins I and II in GH3 rat tumoral prolactin (PRL) cells as revealed by electron microscopic immunocytochemistry. J Histochem Cytochem. 1989 Sep;37(9):1329–1336. [PubMed]
  • Vaux D, Tooze J, Fuller S. Identification by anti-idiotype antibodies of an intracellular membrane protein that recognizes a mammalian endoplasmic reticulum retention signal. Nature. 1990 Jun 7;345(6275):495–502. [PubMed]
  • Wiedenmann B, Huttner WB. Synaptophysin and chromogranins/secretogranins--widespread constituents of distinct types of neuroendocrine vesicles and new tools in tumor diagnosis. Virchows Arch B Cell Pathol Incl Mol Pathol. 1989;58(2):95–121. [PubMed]
  • Yoo SH, Albanesi JP. Ca2(+)-induced conformational change and aggregation of chromogranin A. J Biol Chem. 1990 Aug 25;265(24):14414–14421. [PubMed]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...