Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1987 May;6(5):1203–1211. doi: 10.1002/j.1460-2075.1987.tb02355.x

The primary structure of human secretogranin I (chromogranin B): comparison with chromogranin A reveals homologous terminal domains and a large intervening variable region.

U M Benedum, A Lamouroux, D S Konecki, P Rosa, A Hille, P A Baeuerle, R Frank, F Lottspeich, J Mallet, W B Huttner
PMCID: PMC553920  PMID: 3608978

Abstract

We have determined and analyzed the primary structure of human secretogranin I (chromogranin B), a tyrosine-sulfated secretory protein found in a wide variety of peptidergic endocrine cells. A 2.5-kb cDNA clone, hybridizing to an mRNA of similar length, was isolated from a cDNA library of human pheochromocytoma. The identity of the clone was established by comparison of its deduced amino acid sequence with N-terminal and several internal secretogranin I sequences as well as by immunoprecipitation of the protein produced by in vitro transcription-translation of the cloned cDNA. Secretogranin I is a 657 amino acid long polypeptide of 76 kd and is preceded by a cleaved N-terminal signal peptide of 20 residues. Comparison of the predicted amino acid sequence of human secretogranin I with that of bovine chromogranin A reveals significant homologies near the N termini and at the C termini. The N-terminal homologous domains contain the only two cysteine residues of both proteins and form disulfide-stabilized loop structures. The sequences between the homologous terminal domains in both proteins differ but are characterized by a remarkable hydrophilicity, an abundance of acidic amino acids and potential dibasic cleavage sites for the generation of smaller, perhaps hormone-like, peptides.

Full text

PDF
1203

Images in this article

1204Fig. 1.
on p.1204
1206Fig. 3.
on p.1206
1207Fig. 4.
on p.1207
1209Fig. 7.
on p.1209

Selected References

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

  1. Argos P. A sensitive procedure to compare amino acid sequences. J Mol Biol. 1987 Jan 20;193(2):385–396. doi: 10.1016/0022-2836(87)90226-9. [DOI] [PubMed] [Google Scholar]
  2. Argos P. Evidence for a repeating domain in type I restriction enzymes. EMBO J. 1985 May;4(5):1351–1355. doi: 10.1002/j.1460-2075.1985.tb03784.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benedum U. M., Baeuerle P. A., Konecki D. S., Frank R., Powell J., Mallet J., Huttner W. B. 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. doi: 10.1002/j.1460-2075.1986.tb04388.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohn D. V., Elting J. J., Frick M., Elde R. Selective localization of the parathyroid secretory protein-I/adrenal medulla chromogranin A protein family in a wide variety of endocrine cells of the rat. Endocrinology. 1984 Jun;114(6):1963–1974. doi: 10.1210/endo-114-6-1963. [DOI] [PubMed] [Google Scholar]
  7. Dobberstein B., Garoff H., Warren G., Robinson P. J. Cell-free synthesis and membrane insertion of mouse H-2Dd histocompatibility antigen and beta 2-microglobulin. Cell. 1979 Aug;17(4):759–769. doi: 10.1016/0092-8674(79)90316-7. [DOI] [PubMed] [Google Scholar]
  8. Docherty K., Steiner D. F. Post-translational proteolysis in polypeptide hormone biosynthesis. Annu Rev Physiol. 1982;44:625–638. doi: 10.1146/annurev.ph.44.030182.003205. [DOI] [PubMed] [Google Scholar]
  9. Eiden L. E. Is chromogranin a prohormone? Nature. 1987 Jan 22;325(6102):301–301. doi: 10.1038/325301a0. [DOI] [PubMed] [Google Scholar]
  10. Falkensammer G., Fischer-Colbrie R., Richter K., Winkler H. Cell-free and cellular synthesis of chromogranin A and B of bovine adrenal medulla. Neuroscience. 1985 Feb;14(2):735–746. doi: 10.1016/0306-4522(85)90323-9. [DOI] [PubMed] [Google Scholar]
  11. Falkensammer G., Fischer-Colbrie R., Winkler H. Biogenesis of chromaffin granules: incorporation of sulfate into chromogranin B and into a proteoglycan. J Neurochem. 1985 Nov;45(5):1475–1480. doi: 10.1111/j.1471-4159.1985.tb07215.x. [DOI] [PubMed] [Google Scholar]
  12. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  13. Fischer-Colbrie R., Diez-Guerra J., Emson P. C., Winkler H. Bovine chromaffin granules: immunological studies with antisera against neuropeptide Y, [Met]enkephalin and bombesin. Neuroscience. 1986 May;18(1):167–174. doi: 10.1016/0306-4522(86)90185-5. [DOI] [PubMed] [Google Scholar]
  14. Fischer-Colbrie R., Lassmann H., Hagn C., Winkler H. Immunological studies on the distribution of chromogranin A and B in endocrine and nervous tissues. Neuroscience. 1985 Nov;16(3):547–555. doi: 10.1016/0306-4522(85)90191-5. [DOI] [PubMed] [Google Scholar]
  15. Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
  16. Hagn C., Schmid K. W., Fischer-Colbrie R., Winkler H. Chromogranin A, B, and C in human adrenal medulla and endocrine tissues. Lab Invest. 1986 Oct;55(4):405–411. [PubMed] [Google Scholar]
  17. Hille A., Rosa P., Huttner W. B. Tyrosine sulfation: a post-translational modification of proteins destined for secretion? FEBS Lett. 1984 Nov 5;177(1):129–134. doi: 10.1016/0014-5793(84)80996-5. [DOI] [PubMed] [Google Scholar]
  18. Hortsch M., Meyer D. I. Transfer of secretory proteins through the membrane of the endoplasmic reticulum. Int Rev Cytol. 1986;102:215–242. doi: 10.1016/s0074-7696(08)61276-0. [DOI] [PubMed] [Google Scholar]
  19. Huttner W. B., Benedum U. M. Chromogranin A and pancreastatin. Nature. 1987 Jan 22;325(6102):305–305. doi: 10.1038/325305b0. [DOI] [PubMed] [Google Scholar]
  20. Hörtnagl H., Lochs H., Winkler H. Immunological studies on the acidic chromogranins and on dopamine beta-hydroxylase (EC 1.14.2.1) of bovine chromaffin granules. J Neurochem. 1974 Jan;22(1):197–199. doi: 10.1111/j.1471-4159.1974.tb12201.x. [DOI] [PubMed] [Google Scholar]
  21. Iacangelo A., Affolter H. U., Eiden L. E., 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. doi: 10.1038/323082a0. [DOI] [PubMed] [Google Scholar]
  22. Kelly R. B. Pathways of protein secretion in eukaryotes. Science. 1985 Oct 4;230(4721):25–32. doi: 10.1126/science.2994224. [DOI] [PubMed] [Google Scholar]
  23. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lassmann H., Hagn C., Fischer-Colbrie R., Winkler H. Presence of chromogranin A, B and C in bovine endocrine and nervous tissues: a comparative immunohistochemical study. Histochem J. 1986 Jul;18(7):380–386. doi: 10.1007/BF01675219. [DOI] [PubMed] [Google Scholar]
  26. Lee R. W., Huttner W. B. (Glu62, Ala30, Tyr8)n serves as high-affinity substrate for tyrosylprotein sulfotransferase: a Golgi enzyme. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6143–6147. doi: 10.1073/pnas.82.18.6143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lee R. W., Huttner W. B. Tyrosine-O-sulfated proteins of PC12 pheochromocytoma cells and their sulfation by a tyrosylprotein sulfotransferase. J Biol Chem. 1983 Sep 25;258(18):11326–11334. [PubMed] [Google Scholar]
  28. Lloyd R. V., Wilson B. S. Specific endocrine tissue marker defined by a monoclonal antibody. Science. 1983 Nov 11;222(4624):628–630. doi: 10.1126/science.6635661. [DOI] [PubMed] [Google Scholar]
  29. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  30. O'Connor D. T., Burton D., Deftos L. J. Chromogranin A: immunohistology reveals its universal occurrence in normal polypeptide hormone producing endocrine glands. Life Sci. 1983 Oct 24;33(17):1657–1663. doi: 10.1016/0024-3205(83)90721-x. [DOI] [PubMed] [Google Scholar]
  31. O'Connor D. T., Frigon R. P. Chromogranin A, the major catecholamine storage vesicle soluble protein. Multiple size forms, subcellular storage, and regional distribution in chromaffin and nervous tissue elucidated by radioimmunoassay. J Biol Chem. 1984 Mar 10;259(5):3237–3247. [PubMed] [Google Scholar]
  32. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  33. Rindi G., Buffa R., Sessa F., Tortora O., Solcia E. Chromogranin A, B and C immunoreactivities of mammalian endocrine cells. Distribution, distinction from costored hormones/prohormones and relationship with the argyrophil component of secretory granules. Histochemistry. 1986;85(1):19–28. doi: 10.1007/BF00508649. [DOI] [PubMed] [Google Scholar]
  34. Rosa P., Fumagalli G., Zanini A., Huttner W. B. The major tyrosine-sulfated protein of the bovine anterior pituitary is a secretory protein present in gonadotrophs, thyrotrophs, mammotrophs, and corticotrophs. J Cell Biol. 1985 Mar;100(3):928–937. doi: 10.1083/jcb.100.3.928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rosa P., Hille A., Lee R. W., Zanini A., De Camilli P., Huttner W. B. 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. doi: 10.1083/jcb.101.5.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Settleman J., Fonseca R., Nolan J., Angeletti R. H. Relationship of multiple forms of chromogranin. J Biol Chem. 1985 Feb 10;260(3):1645–1651. [PubMed] [Google Scholar]
  39. Sinha N. D., Biernat J., McManus J., Köster H. Polymer support oligonucleotide synthesis XVIII: use of beta-cyanoethyl-N,N-dialkylamino-/N-morpholino phosphoramidite of deoxynucleosides for the synthesis of DNA fragments simplifying deprotection and isolation of the final product. Nucleic Acids Res. 1984 Jun 11;12(11):4539–4557. doi: 10.1093/nar/12.11.4539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Somogyi P., Hodgson A. J., DePotter R. W., Fischer-Colbrie R., Schober M., Winkler H., Chubb I. W. Chromogranin immunoreactivity in the central nervous system. Immunochemical characterisation, distribution and relationship to catecholamine and enkephalin pathways. Brain Res. 1984 Dec;320(2-3):193–230. doi: 10.1016/0165-0173(84)90007-9. [DOI] [PubMed] [Google Scholar]
  41. Stueber D., Ibrahimi I., Cutler D., Dobberstein B., Bujard H. A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences. EMBO J. 1984 Dec 20;3(13):3143–3148. doi: 10.1002/j.1460-2075.1984.tb02271.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tatemoto K., Efendić S., Mutt V., Makk G., Feistner G. J., Barchas J. D. Pancreastatin, a novel pancreatic peptide that inhibits insulin secretion. Nature. 1986 Dec 4;324(6096):476–478. doi: 10.1038/324476a0. [DOI] [PubMed] [Google Scholar]
  43. Trifaró J. M., Duerr A. C. Isolation and characterization of a Golgi-rich fraction from the adrenal medulla. Biochim Biophys Acta. 1976 Jan 14;421(1):153–167. doi: 10.1016/0304-4165(76)90179-3. [DOI] [PubMed] [Google Scholar]
  44. Warren G., Dobberstein B. Protein transfer across microsomal membranes reassembled from separated membrane components. Nature. 1978 Jun 15;273(5663):569–571. doi: 10.1038/273569a0. [DOI] [PubMed] [Google Scholar]
  45. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES