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Molecular characterization and tissue expression of carboxypeptidase H (CPH) gene in flounder (Paralichthys olivaceus)

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Abstract

Carboxypeptidase H (CPH) is one of several enzymes required for the processing of peptide hormone precursors and removal of basic amino acids from intermediates during protein biosynthesis. The CPH gene was isolated from a flounder (Paralichthys olivaceus) brain cDNA library and was found to consist of 1,590 bp encoding 455 amino acids residues. The nucleotide sequence of the CPH gene was compared with those of other species, including zebrafish and mouse, and turned out to be conserved during evolution. Expression of CPH mRNA was confirmed by both Reverse transcription polymerase Chain Reaction (RT-PCR) and Northern blot analysis. A recombinant protein of approximately 50 kDa, which corresponds to CPH, was functionally expressed in E. coli. Biochemical studies on the expressed protein were performed. We report here the molecular characteristics and tissue expression of newly identified CPH cDNA from flounder (P. olivaceus).

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References

  1. Yao, J., D. Zadworny, U. Kuhnlein, and J. F. Hayes (1995) Molecular cloning of a bovine ornithine decarboxylase cDNA and its use in the detection of restriction fragment length polymorphisms in Holsteins. Genome 38: 325–331.

    Article  CAS  Google Scholar 

  2. Docherty, K. and D. F. Steiner (1982) Post-translational proteolysis in polypeptide hormone biosynthesis. Annu. Rev. Physiol. 44: 625–638.

    Article  CAS  Google Scholar 

  3. Steiner, D. F. (1991) The biosynthesis of biologically active peptides, a perspective. pp. 1–15. In: L. D. Fricker (ed.). Peptide Biosynthesis and Processing. CRC Press, Florida, USA.

    Google Scholar 

  4. Webb, E. C. (1986) Enzyme nomenclature of the International union of biochemistry. Eur.J.Biochem. 157: 1–26.

    Article  Google Scholar 

  5. Fricker, L. D. (1988) Carboxypeptidase E. Annu. Rev. Physiol. 50: 309–321.

    Article  CAS  Google Scholar 

  6. Fricker, L. D. and S. H. Snyder (1982) Enkephalin convertase: Purification and characterization of specific enkephalin-synthesizing carboxypeptidase localized to adrenal chromaffin granules. Proc. Natl. Acad. Sci. 79: 3886–3890.

    Article  CAS  Google Scholar 

  7. Fricker, L. D. and S. H. Snyder (1983) Purification and characterization of enkephalin convertase, enkephalin-synthesizing carboxypeptidase. J. Biol. Chem. 258: 10950–10955.

    CAS  Google Scholar 

  8. Hook, V. Y., Eiden, L. E., and M. J. Brownstein (1982) A carboxypeptidase processing enzyme for enkephalin precursors. Nature. 295: 341–342.

    Article  CAS  Google Scholar 

  9. Hook, V. Y., and Y. P. Loh. (1984) Carboxypeptidase B-like converting enzyme activity in secretory granules of rat pituitary. Proc. Natl. Acad. Sci. U.S.A. 81: 2776–2787.

    Article  CAS  Google Scholar 

  10. Davidson, H. W. and J. C. Hutton (1987) The insulin-secretorygranule carboxypeptidase H, purification and demonstration of involvement in proinsulin processing. Biochem. J. 245: 575–582.

    CAS  Google Scholar 

  11. Schwartz, T. W. (1990) The processing of peptide precursors. pp. 153–205. In: Hiroshi Okamoto (ed.). Molecular Biology of Islets of Langerhans. Cambridge University Press, Cambridge, UK.

    Chapter  Google Scholar 

  12. Burgess, T. L. and R. B. Kelly (1987) Constitutive and regulated secretion of proteins. Annu. Rev. Cell. Biol. 3: 243–293.

    Article  CAS  Google Scholar 

  13. Laslop, A., C. R. Fischer, V. Hook, D. A. Obendorf, and Winkler, H. (1986) Identification of two gycoproteins of chromaffin granules as the carboxypeptidase H. Neurosci. Lett. 72: 300–304.

    Article  CAS  Google Scholar 

  14. Fricker, L. D. and E. Herbert (1988) Comparison of a carboxypeptidase E-like enzyme in human, bovine, mouse, Xenopus, shark and Aplysia neural tissue. Brain. Res. 453: 281–286.

    Article  CAS  Google Scholar 

  15. Fricker, L, D., B. Das, and R. H. Angeletti (1990) Identification of the pH dependent membrane anchor of carboxypeptidase E (EC 3.4.17.10). J. Biol. Chem. 265: 2476–2482.

    CAS  Google Scholar 

  16. Naggert, J. K., L. D. Fricker, O. Varlamov, P. M. Nishina, Y. Rouille, D. F. Steiner, R. J. Carroll, B. J. Paigen, and E. H. Leiter (1995) Hyperproinsulinaemia in obese fat/fat mice associated with a carboxypeptidase E mutation which reduces enzyme activity. Nat. Genet. 10: 135–142.

    Article  CAS  Google Scholar 

  17. Fricker, L. D., C. J. Evans, F. S. Esch, and E. Herbert (1986) Cloning and sequence analysis of cDNA for bovine carboxypeptidase E. Nature 323: 461–464.

    Article  CAS  Google Scholar 

  18. Fricker, L. D., J. P. Adelman, J. Douglass, R. C. Thompson, R. P. von Strandmann, and J. Hutton (1989) Isolation and sequence analysis of cDNA for rat carboxypeptidase E [EC 3.4.17.10], a neuropeptide processing enzyme. Mol. Endocrinol. 3: 666–673.

    Article  CAS  Google Scholar 

  19. Manser, E., D. Fernandez, L. Loo, P. Y. Goh, C. Monfries, C. Hall, and L. Lim (1990) Human carboxypeptidase E. Isolation and characterization of the cDNA, sequence conservation, expression and processing in vitro. Biochem. J. 267: 517–525.

    CAS  Google Scholar 

  20. Roth, W. W., R. B. Mackin, J. Spiess, R. H. Goodman, and B. D. Noe (1991) Primary structure and tissue distribution of angler fish carboxypeptidase H. Mol. Cell. Endocrinol. 78: 171–178.

    Article  CAS  Google Scholar 

  21. Woronowicz, A., N. X. Cawley, S. Y. Chang, H. Koshimizu, A. W. Phillips, Z. G. Xiong, and Y. P. Loh (2010) Carboxypeptidase E knockout mice exhibit abnormal dendritic arborization and spine morphology in central nervous system neurons. J. Neurosci. Res. 88: 64–72.

    Article  CAS  Google Scholar 

  22. Jung, Y. K., C. J. Kunczt, R. K. Pearson, J. E. Dixon, and L. D. Fricker (1991) Structural characterization of the rat carboxypeptidase-E gene. Mol. Endocrinol. 5: 1257–1268.

    Article  CAS  Google Scholar 

  23. Klein, S. L., R. L. Strausberg, L. Wagner, J. Pontius, S. W. Clifton, and P. Richardson (2002) Genetic and genomic tools for Xenopus research: The NIH Xenopus initiative. Dev. Dyn. 225: 384–391.

    Article  CAS  Google Scholar 

  24. Zhang, X., J. Zhu, Y. P. Loh, and L. R. Berghman (2009) Carboxypeptidase E, an essential element of the regulated secretory pathway, is expressed and partially co-localized with chromogranin A in chicken thymus. Cell Tissue Res. 337: 371–379.

    Article  CAS  Google Scholar 

  25. Bernasconi, P. (1994) Molecular cloning of a Drosophila melanogaster gene coding for an homologue of human carboxypeptidase E. Arch. Insect Biochem. Physiol. 27: 169–178.

    Article  CAS  Google Scholar 

  26. Christie, D. L. and D. J. Palmer (1990) Identification and characterization of glycoproteins after extraction of bovine chromaffin-granule membranes with lithium di-iodosalicylate. Purification of glycoprotein II from the soluble fraction. Biochem. J. 270: 57–61.

    CAS  Google Scholar 

  27. Vihtelic, T. S., J. M. Fadool, J. Gao, K. A. Thornton, D. R. Hyde, and G. Wistow (2005) Expressed sequence tag analysis of zebrafish eye tissues for NEI Bank. Mol. Vis. 11: 1083–1100.

    CAS  Google Scholar 

  28. Kim, D. S. and Y. T. Kim (1999) Identification of an embryonic growth factor IGF-II from the central nervous system of the teleost, flounder, and its expressions in adult tissues. J. Microbiol. Biotech. 9: 113–118.

    CAS  Google Scholar 

  29. Cho, J. J., B. K. Sung, J. H. Lee, J. K. Chung, T. J. Choi, and Y. T. Kim (2001) cDNA for an immune response gene encoding low molecular weight polypeptide from flounder, Paralichthys olivaceus. Mol. Cells 11: 226–230.

    CAS  Google Scholar 

  30. Lee, E. Y., H. H. Park, Y. T. Kim, and T. J. Choi (2001) Cloning and sequence analysis of the interleukin-8 gene from flounder (Paralithys olivaceus). Gene 274: 237–243.

    Article  CAS  Google Scholar 

  31. Kim, Y. T. and C. C. Richardson (1994) Acidic carboxyl-terminal domain of gene 2.5 protein of bacteriophage T7 is essential for protein-protein interactions. J. Biol. Chem. 269: 5270–5278.

    CAS  Google Scholar 

  32. Kim, Y. T., Y. H. Song, and J. E. Churchich (1997) Recombinant brain 4-aminobutyrate aminotransferases: Overexpression, purification, and identification of Lys-330 at the active site. Biochim. Biophys. Acta 1337: 248–256.

    Article  CAS  Google Scholar 

  33. Cho, J. J., J. H. Lee, S. K. Kim, T. J. Choi, and Y. T. Kim (1999) cDNA encoding nm23/NDP kinase gene from Korean tiger shark, Scyliorhinus torazame. Mar. Biotechnol. 1: 131–136.

    Article  CAS  Google Scholar 

  34. Van de Peer, Y. and R. De Wachter (1994) TREECON for Windows: A software package for the construction and drawing of evolutionary trees for the microsoft Windows environment. Comput. Appl. Biosci. 10: 569–570.

    Google Scholar 

  35. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  CAS  Google Scholar 

  36. Gershoni, J. M. and G. E. Palade (1983) Protein blotting: Principles and application. Anal. Biochem. 131: 1–15.

    Article  CAS  Google Scholar 

  37. Folk, J. E., K. A. Piez, W. R. Carroll, and J. A. Gladner (1960) Carboxy-peptidase B. 4. Purification and characterization of the porcine enzyme. J. Biol. Chem. 235: 2272–2277.

    CAS  Google Scholar 

  38. Aloy, P., V. Companys, J. Vendrell, F. X. Aviles, L. D. Fricker, M. Coll, and F. Xavier Gomis-Ruth (2001) The crystal structure of the inhibitor-complexed carboxypeptidase D domain II and the modeling of regulatory carboxypeptidases. J. Biol. Chem. 276: 16177–16184.

    Article  CAS  Google Scholar 

  39. Zuhlke, H., K.-D. Kohnert, H. Jahr, S. Schmidt, H. Kirschke, and D. F. Steiner (1977) Proteolytic and transhydrogenolytic activities in isolated pancreatic islets of rats. Acta Biol. Med. Ger. 36: 1695–1703.

    CAS  Google Scholar 

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  1. Department of Microbiology, Pukyong National University, Busan, 608-737, Korea

    Yong Bae Seo, Heeju Hwang & Young Tae Kim

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  1. Yong Bae Seo
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  2. Heeju Hwang
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Correspondence to Young Tae Kim.

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Seo, Y.B., Hwang, H. & Kim, Y.T. Molecular characterization and tissue expression of carboxypeptidase H (CPH) gene in flounder (Paralichthys olivaceus). Biotechnol Bioproc E 16, 374–382 (2011). https://doi.org/10.1007/s12257-010-0278-6

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  • DOI: https://doi.org/10.1007/s12257-010-0278-6

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