Logo of embojLink to Publisher's site
EMBO J. 1999 Feb 15; 18(4): 1059–1070.
PMCID: PMC1171197

The disulfide-bonded loop of chromogranin B mediates membrane binding and directs sorting from the trans-Golgi network to secretory granules.

Abstract

The disulfide-bonded loop of chromogranin B (CgB), a regulated secretory protein with widespread distribution in neuroendocrine cells, is known to be essential for the sorting of CgB from the trans-Golgi network (TGN) to immature secretory granules. Here we show that this loop, when fused to the constitutively secreted protein alpha1-antitrypsin (AT), is sufficient to direct the fusion protein to secretory granules. Importantly, the sorting efficiency of the AT reporter protein bearing two loops (E2/3-AT-E2/3) is much higher compared with that of AT with a single disulfide-bonded loop. In contrast to endogenous CgB, E2/3-AT-E2/3 does not undergo Ca2+/pH-dependent aggregation in the TGN. Furthermore, the disulfide-bonded loop of CgB mediates membrane binding in the TGN and does so with 5-fold higher efficiency if two loops are present on the reporter protein. The latter finding supports the concept that under physiological conditions, aggregates of CgB are the sorted units of cargo which have multiple loops on their surface leading to high membrane binding and sorting efficiency of CgB in the TGN.

Full Text

The Full Text of this article is available as a PDF (566K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aaku-Saraste E, Hellwig A, Huttner WB. Loss of occludin and functional tight junctions, but not ZO-1, during neural tube closure--remodeling of the neuroepithelium prior to neurogenesis. Dev Biol. 1996 Dec 15;180(2):664–679. [PubMed]
  • Arvan P, Castle D. Protein sorting and secretion granule formation in regulated secretory cells. Trends Cell Biol. 1992 Nov;2(11):327–331. [PubMed]
  • Arvan P, Castle D. Sorting and storage during secretory granule biogenesis: looking backward and looking forward. Biochem J. 1998 Jun 15;332(Pt 3):593–610. [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]
  • 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]
  • Brechler V, Chu WN, Baxter JD, Thibault G, Reudelhuber TL. A protease processing site is essential for prorenin sorting to the regulated secretory pathway. J Biol Chem. 1996 Aug 23;271(34):20636–20640. [PubMed]
  • Burgess TL, Kelly RB. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. [PubMed]
  • Castle AM, Stahl LE, Castle JD. A 13-amino acid n-terminal domain of a basic proline-rich protein is necessary for storage in secretory granules and facilitates exit from the endoplasmic reticulum. J Biol Chem. 1992 Jun 25;267(18):13093–13100. [PubMed]
  • Chanat E, Huttner WB. Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network. J Cell Biol. 1991 Dec;115(6):1505–1519. [PMC free article] [PubMed]
  • Chanat E, Weiss U, Huttner WB. The disulfide bond in chromogranin B, which is essential for its sorting to secretory granules, is not required for its aggregation in the trans-Golgi network. FEBS Lett. 1994 Sep 5;351(2):225–230. [PubMed]
  • Chanat E, Weiss U, Huttner WB, Tooze SA. Reduction of the disulfide bond of chromogranin B (secretogranin I) in the trans-Golgi network causes its missorting to the constitutive secretory pathways. EMBO J. 1993 May;12(5):2159–2168. [PMC free article] [PubMed]
  • Chevrier D, Fournier H, Nault C, Zollinger M, Crine P, Boileau G. Targeting of pro-opiomelanocortin to the regulated secretory pathway may involve cooperation between different protein domains. Mol Cell Endocrinol. 1993 Aug;94(2):213–221. [PubMed]
  • Cool DR, Loh YP. Identification of a sorting signal for the regulated secretory pathway at the N-terminus of pro-opiomelanocortin. Biochimie. 1994;76(3-4):265–270. [PubMed]
  • Cool DR, Fenger M, Snell CR, Loh YP. Identification of the sorting signal motif within pro-opiomelanocortin for the regulated secretory pathway. J Biol Chem. 1995 Apr 14;270(15):8723–8729. [PubMed]
  • Cool DR, Normant E, Shen F, Chen HC, Pannell L, Zhang Y, Loh YP. Carboxypeptidase E is a regulated secretory pathway sorting receptor: genetic obliteration leads to endocrine disorders in Cpe(fat) mice. Cell. 1997 Jan 10;88(1):73–83. [PubMed]
  • Corti A, Sanchez LP, Gasparri A, Curnis F, Longhi R, Brandazza A, Siccardi AG, Sidoli A. Production and structure characterisation of recombinant chromogranin A N-terminal fragments (vasostatins) -- evidence of dimer-monomer equilibria. Eur J Biochem. 1997 Sep 15;248(3):692–699. [PubMed]
  • Forss-Petter S, Danielson P, Battenberg E, Bloom F, Sutcliffe JG. Nucleotide sequence and cellular distribution of rat chromogranin B (secretogranin I) mRNA in the neuroendocrine system. J Mol Neurosci. 1989;1(2):63–75. [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]
  • Gorr SU, Darling DS. An N-terminal hydrophobic peak is the sorting signal of regulated secretory proteins. FEBS Lett. 1995 Mar 13;361(1):8–12. [PubMed]
  • Huttner WB, Ohashi M, Kehlenbach RH, Barr FA, Bauerfeind R, Bräunling O, Corbeil D, Hannah M, Pasolli HA, Schmidt A, et al. Biogenesis of neurosecretory vesicles. Cold Spring Harb Symp Quant Biol. 1995;60:315–327. [PubMed]
  • Kelly RB. Pathways of protein secretion in eukaryotes. Science. 1985 Oct 4;230(4721):25–32. [PubMed]
  • Krömer A, Glombik MM, Huttner WB, Gerdes HH. Essential role of the disulfide-bonded loop of chromogranin B for sorting to secretory granules is revealed by expression of a deletion mutant in the absence of endogenous granin synthesis. J Cell Biol. 1998 Mar 23;140(6):1331–1346. [PMC free article] [PubMed]
  • Leitinger B, Brown JL, Spiess M. Tagging secretory and membrane proteins with a tyrosine sulfation site. Tyrosine sulfation precedes galactosylation and sialylation in COS-7 cells. J Biol Chem. 1994 Mar 18;269(11):8115–8121. [PubMed]
  • Natori S, King A, Hellwig A, Weiss U, Iguchi H, Tsuchiya B, Kameya T, Takayanagi R, Nawata H, Huttner WB. Chromogranin B (secretogranin I), a neuroendocrine-regulated secretory protein, is sorted to exocrine secretory granules in transgenic mice. EMBO J. 1998 Jun 15;17(12):3277–3289. [PMC free article] [PubMed]
  • Normant E, Loh YP. Depletion of carboxypeptidase E, a regulated secretory pathway sorting receptor, causes misrouting and constitutive secretion of proinsulin and proenkephalin, but not chromogranin A. Endocrinology. 1998 Apr;139(4):2137–2145. [PubMed]
  • Pimplikar SW, Huttner WB. Chromogranin B (secretogranin I), a secretory protein of the regulated pathway, is also present in a tightly membrane-associated form in PC12 cells. J Biol Chem. 1992 Feb 25;267(6):4110–4118. [PubMed]
  • Pohl TM, Phillips E, Song KY, Gerdes HH, Huttner WB, Rüther U. The organisation of the mouse chromogranin B (secretogranin I) gene. FEBS Lett. 1990 Mar 26;262(2):219–224. [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]
  • Scheiffele P, Peränen J, Simons K. N-glycans as apical sorting signals in epithelial cells. Nature. 1995 Nov 2;378(6552):96–98. [PubMed]
  • Sevarino KA, Stork P. Multiple preprosomatostatin sorting signals mediate secretion via discrete cAMP- and tetradecanoylphorbolacetate-responsive pathways. J Biol Chem. 1991 Oct 5;266(28):18507–18513. [PubMed]
  • Shaw WV, Leslie AG. Chloramphenicol acetyltransferase. Annu Rev Biophys Biophys Chem. 1991;20:363–386. [PubMed]
  • Stoller TJ, Shields D. The propeptide of preprosomatostatin mediates intracellular transport and secretion of alpha-globin from mammalian cells. J Cell Biol. 1989 May;108(5):1647–1655. [PMC free article] [PubMed]
  • Tam WW, Andreasson KI, Loh YP. The amino-terminal sequence of pro-opiomelanocortin directs intracellular targeting to the regulated secretory pathway. Eur J Cell Biol. 1993 Dec;62(2):294–306. [PubMed]
  • Thiele C, Huttner WB. The disulfide-bonded loop of chromogranins, which is essential for sorting to secretory granules, mediates homodimerization. J Biol Chem. 1998 Jan 9;273(2):1223–1231. [PubMed]
  • Thiele C, Gerdes HH, Huttner WB. Protein secretion: puzzling receptors. Curr Biol. 1997 Aug 1;7(8):R496–R500. [PubMed]
  • Tong PY, Gregory W, Kornfeld S. Ligand interactions of the cation-independent mannose 6-phosphate receptor. The stoichiometry of mannose 6-phosphate binding. J Biol Chem. 1989 May 15;264(14):7962–7969. [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]
  • Varlamov O, Fricker LD. The C-terminal region of carboxypeptidase E involved in membrane binding is distinct from the region involved with intracellular routing. J Biol Chem. 1996 Mar 15;271(11):6077–6083. [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem chemical compound records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records. Multiple substance records may contribute to the PubChem compound record.
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...