Sequence Analysis of the Carboxypeptidase E Precursor

  • Lloyd D. Fricker
Part of the Biochemical Endocrinology book series (BIOEND)


With the use of recombinant DNA technology, much has recently been learned about precursors of proteins and bioactive peptides. Sequencing of cDNA clones has revealed that many proteins are originally produced as larger precursors. These protein precursors must be post-translationally processed into the active protein. One of the best studied precursor sequences is the ‘signal peptide’, which is typically a hydrophobic 20–25 amino acid peptide located on the N-terminus of the protein precursor (Von Heijne, 1985). The signal peptide is usually removed by a signal peptidase located in the endoplasmic reticulum. Most proteins that are secreted or localized to subcellular organelles initially contain signal peptides, which directs the translocation of newly synthesized proteins into the rough endoplasmic reticulum. In addition to signal peptides, many proteins contain other precursor sequences. Examples of proteins that are produced from larger precursors (containing more than a signal peptide) include receptors, such as the insulin receptor (Ebina et al, 1985), enzymes, such as carboxypeptidase A (Quinto et al, 1982) and prothrombin (Degen et al, 1983), and numerous peptide hormones and neurotransmitters (Docherty and Steiner, 1982).


Signal Peptide Secretory Granule Basic Amino Acid Chromaffin Granule Signal Peptide Cleavage Site 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Belt, K.T., Carroll, M.C., and Porter, R.R., 1985, The structural basis of the multiple forms of human complement component C4 Cell 36:907.CrossRefGoogle Scholar
  2. Bentley, A.K., Rees, D.J.G., Rizza, C., and Brownlee, G.G., 1986, Defective propeptide processing of blood clotting factor IX caused by mutation of arginine to glutamine at position -4 Cell 45:343.PubMedCrossRefGoogle Scholar
  3. Cromlish, J.A., Seidah, N.G., and Chretien, M., 1986, Selective cleavage of human ACTH, beta-lipotropin, and the N-terminal glycopeptide at pairs of basic residues by IRCM-serine protease 1 J. Biol Chem. 261:868.Google Scholar
  4. de Bruijn, M.H.L., and Fey, G.H., Human complement component C3: cDNA coding sequence and derived primary structure Proc. Natl. Acad. Sci. USA 82:708Google Scholar
  5. Degen, S.J.F., MacGillivray, R.T.A., and Davie, E.W., 1983, Characterization of the complementary deoxyribonucleic acid and gene coding for human prothrombin Biochem 22:2087.CrossRefGoogle Scholar
  6. Docherty, K., and Hutton, J.C., 1983, Carboxypeptidase activity in the insulin secretory granule FEBS Letters 162:137.PubMedCrossRefGoogle Scholar
  7. Docherty, K., and Steiner, D.F., 1982, Post-translational proteolysis in polypeptide hormone biosynthesis Ann. Rev. Physiol. 44:625.CrossRefGoogle Scholar
  8. Ebina, Y., Ellis, L., Jarnagin, K., Edery, M., Graf, L., Clauser, E., Ou, J., Masiarz, F., Kan, Y.W., Goldfine, I.D., Roth, R.A., and Rutter, W.J., 1985, The human insulin receptor cDNA: The structural basis for hormone-activated transmembrane signalling Cell 40:747.PubMedCrossRefGoogle Scholar
  9. Fischer-Colbrie, R., and Frischenschlager, I., 1985, Immunological characterization of secretory proteins of chromaffin granules: Chromogranins A, chromogranins B, and enkephalin-containing peptides J. Neurochem. 44:1854.PubMedCrossRefGoogle Scholar
  10. Folk, J.E., 1971, Carboxypeptidase B in: Enzymes (Boyer, P.D. eds.) 3rd Edition, page 57 Academic Press, New York.Google Scholar
  11. Fricker, L.D., and Herbert, E., 1987, Molecular biology of carboxypeptidase E (enkephalin convertase), a neuropeptide synthesizing enzyme Annals NY Acad. Sci. 493:391.CrossRefGoogle Scholar
  12. Fricker, L.D., and Snyder, S.H., 1982, Enkephalin convertase: Purification and characterization of a specific enkephalin-synthesizing carboxypeptidase localized to adrenal chromaffin granules Proc. Natl. Acad. Sci. USA 79:3886.CrossRefGoogle Scholar
  13. Fricker, L.D., and Snyder, S.H., 1983, Purification and characterization of enkephalin convertase, an enkephalin-synthesizing carboxypeptidase J. Biol. Chem. 258:10950.PubMedGoogle Scholar
  14. Fricker, L.D., Evans, C.J., Esch, F.S., and and Herbert, E., 1986, Cloning and sequence analysis of cDNA for bovine carboxypeptidase E Nature 323:461.PubMedCrossRefGoogle Scholar
  15. Hindley, J., 1983, DNA sequencing in: Laboratory methods in biochemistry and molecular biology (Work, T.S. Burdon, R.H. eds.) page 121 Elsevier Biomedical Press, Amsterdam.Google Scholar
  16. Hook, V.Y.H., and Loh, Y.P., 1984, Carboxypeptidase B-like converting enzyme activity in secretory granules of rat pituitary Proc. Natl. Acad. Sci. USA 81:2776.CrossRefGoogle Scholar
  17. Iacangelo, A., Affolter, H.U., Eiden, L.E., Herbert, E., and Grimes, M., 1986, Bovine chromogranin A sequence and distribution of its messenger RNA in endocrine tissues Nature 323:82.PubMedCrossRefGoogle Scholar
  18. Johnson, L.M., Bankàitis, V.A., and Emr, S.D., 1987, Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease Cell 48:875.PubMedCrossRefGoogle Scholar
  19. Kaiser, E.T., and Kezdy, F.J., 1983, Secondary structures of proteins and peptides in amphiphilic environments (a review) Proc. Natl. Acad. Sci. USA 80:1137.CrossRefGoogle Scholar
  20. Kanmera, T., and Chaiken, I.M., 1985, Pituitary enzyme conversion of putative synthetic oxytocin precursor intermediates J. Biol. Chem. 260:10118.PubMedGoogle Scholar
  21. Kozak, M., 1984, Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs Nuc. Acids Res. 12:857.CrossRefGoogle Scholar
  22. Lindberg, I., Yang, H.Y.T., and Costa, E., 1984, Further characterization of an enkephalin-generating enzyme from adrenal medullary chromaffin granules J. Neurochem. 42:1411.PubMedCrossRefGoogle Scholar
  23. Loh, Y.P., Brownstein, M.J., and Gainer, H., 1984, Proteolysis in neuropeptide processing and other neural functions Ann. Rev. Neurosci. 7:189.PubMedCrossRefGoogle Scholar
  24. Okayama, H., and Berg, P., 1982, High-efficiency cloning of full-length cDNA Molec. Cell. Biol. 2:161.Google Scholar
  25. Plummer, T.H., Jr., and Erdos, E.G., 1981, Human plasma carboxypeptidase N Meth. Enzymol. 80:442.CrossRefGoogle Scholar
  26. Quinto, C., Quiroga, M., Swain, W.F., Nikovits, W.C., Standring, D.N., Pictet, R.L., Valenzuela, P., and Rutter, W.J., 1982, Rat preprocarboxypeptidase Ä: cDNA sequence and preliminary characterization of the gene Proc. Natl. Acad. Sci. USA 79:31.CrossRefGoogle Scholar
  27. Strittmatter, S.M., Lynch, D.R., and Snyder, S.H., 1984, [3H]Guanidinoethylmercaptosuccinic acid binding to tissue homogenates J. Biol. Chem. 259:11812.Google Scholar
  28. Supattapone, S., Fricker, L.D., and Snyder, S.H., 1984, Purification and characterization of a membrane-bound enkephalin-forming carboxypeptidase, “enkephalin convertase” J. Neurochem. 42:1017.PubMedCrossRefGoogle Scholar
  29. Valls, L.A., Hunter, C.P., Rothman, J.H., and Stevens, T.H., 1987, Protein sorting in yeast: The localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide Cell 48:887.PubMedCrossRefGoogle Scholar
  30. Von Heijne, G., 1985, Structural and thermodynamic aspects of the transfer of proteins into and across membranes Curr. Top. Memb. Tranport 24:151.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Lloyd D. Fricker
    • 1
  1. 1.Department of Molecular PharmacologyAlbert Einstein College of MedicineBronxUSA

Personalised recommendations