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Fish Physiology and Biochemistry

, Volume 44, Issue 4, pp 1215–1222 | Cite as

Preparation of biologically active monomeric recombinant zebrafish (Danio rerio) and rainbow trout (Oncorhynchus mykiss) recombinant growth hormones

  • Ewa Ocłoń
  • Gili Solomon
  • Zvi Hayouka
  • Arieh GertlerEmail author
Article

Abstract

Fish growth hormones (GHs) play an important role in regulating growth, metabolism, reproduction, osmoregulation, and immunity and have thus garnered attention for their application in aquaculture. Zebrafish GH (zGH) cDNA or rainbow trout GH (rtGH) cDNA was cloned into the pMon3401 vector, expressed in MON105-competent Escherichia coli and purified to homogeneity. Their biological activity was evidenced by their ability to interact with ovine GH receptor extracellular domain and stimulate GH receptor-mediated proliferation in FDC-P1-3B9 cells stably transfected with rabbit GH receptor. The relative affinity of zGH and rtGH, estimated by IC50, was about 38-fold and 512-fold lower, respectively, than ovine GH. This is likely the reason for the low biological activity in cells with rabbit GH receptor, ~ 36-fold lower for zGH and ~ 107-fold lower for rtGH than for human GH. This was not due to improper refolding, as evidenced by circular dichroism analysis. Predicting the activity of fish GHs is problematic as there is no one single optimal in vitro bioassay; heterologous assays may be ambiguous, and only homologous assays are suitable for measuring activity.

Keywords

Growth hormone Fish Recombinant Zebra fish Rainbow trout 

Notes

Acknowledgments

We thank Dr. Mike Waters for the gift of PDC-P1-3B9 cells.

References

  1. Canosa LF, Chang JP, Peter RE (2007) Neuroendocrine control of growth hormone in fish. Gen Comp Endocrinol 151:1–26CrossRefPubMedGoogle Scholar
  2. Chan YH, Cheng CHK, Chan KM (2003) Recombinant goldfish growth hormones (gfGH-I and -II) expressed in Escherichia coli have similar biological activities. Comp Biochem Physiol A Mol Integ Physiol 135:613–624CrossRefGoogle Scholar
  3. Chung WJ, Huang CL, Gong HY, Ou TY, Hsu JL, Hu SY (2015) Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture. Protein Expr Purif 110:79–88CrossRefPubMedGoogle Scholar
  4. Herrington J, Carter-Su C (2001) Signaling pathways activated by the growth hormone receptor. Trends Endocrinol Metab 12:252–257CrossRefPubMedGoogle Scholar
  5. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefPubMedGoogle Scholar
  6. Moriyama S, Yamamoto H, Sugimoto S, Abe T, Hirano T, Kawauchi H (1993) Oral administration of recombinant salmon growth hormone to rainbow trout, Oncorhynchus mykiss. Aquaculture 112:99–106CrossRefGoogle Scholar
  7. Niv-Spector L, Gonen-Berger D, Gourdou I, Biener E, Gussakovsky EE, Benomar Y, Ramanujan KV, Taouis M, Herman B, Callebaut I, Djiane J, Gertler A (2005) Identification of the hydrophobic strand in the A-B loop of leptin as major binding site III: implications for large-scale preparation of potent recombinant human and ovine leptin antagonists. Biochem J 391:221–230CrossRefPubMedPubMedCentralGoogle Scholar
  8. Paduel A, Chapnik-Cohen N, Gertler A, Elizur A (1999) Preparation and characterization of recombinant dolphin fish (Coryphaena hippurus) growth hormone. Protein Expr Purif 16:417–423CrossRefPubMedGoogle Scholar
  9. Poen S, Pornbanlualap S (2013) Growth hormone from striped catfish (Pangasianodon hypophthalmus): genomic organization, recombinant expression and biological activity. Gene 518:316–324CrossRefPubMedGoogle Scholar
  10. Raver N, Vardy E, Livnah O, Devos R, Gertler A (2002) Comparison of R128Q mutations in human, ovine, and chicken leptins. Gen Comp Endocrinol 126:52–58CrossRefPubMedGoogle Scholar
  11. Santos CRA, Cavaco JEB, Ingleton PM, Power DM (2003) Developmental ontogeny of prolactin and prolactin receptor in the sea bream (Sparus aurata). Gen Comp Endocrinol 132:304–314CrossRefPubMedGoogle Scholar
  12. Solomon G, Reicher S, Gussakovsky EE, Jomain JB, Gertler A (2006) Large-scale preparation and in vitro characterization of biologically active human placental (20 and 22K) and pituitary (20K) growth hormones: placental growth hormones have no lactogenic activity in humans. Growth Hormon IGF Res 16:297–307CrossRefGoogle Scholar
  13. Sreerama N, Woody RW (2000) Estimation of protein secondary structure from circular dichroism spectra: comparison of CONTIN, SELCON and CDSSTR methods with an expanded reference set. Anal Biochem 287:252–260CrossRefPubMedGoogle Scholar
  14. van den Eijnden MJ, Lahaye LL, Strous GJ (2006) Disulfide bonds determine growth hormone receptor folding, dimerisation and ligand binding. J Cell Sci 119:3078–3086CrossRefPubMedGoogle Scholar
  15. Whitmore L, Wallace BA (2004) DICHROWEB: an online server for protein secondary structure analyses from circular dichroism spectroscopic data. Nucleic Acids Res 32:668–673CrossRefGoogle Scholar
  16. Yada T (2007) Growth hormone and fish immune system. Gen Comp Endocrinol 152:353–358CrossRefPubMedGoogle Scholar
  17. Zhou H, Ko WKW, Ho WKK, Stojilkovic SS, Wong AOL (2004) Novel aspects of growth hormone (GH) autoregulation: GH induce GH gene expression in grass carp pituitary cell through autocrine/paracrine mechanisms. Endocrinology 145:4615–4628CrossRefPubMedGoogle Scholar
  18. Zhu T, Goh EL, Graichen R, Ling L, Lobie PE (2001) Signal transduction via the growth hormone receptor. Cell Signal 13:599–616CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Ewa Ocłoń
    • 1
    • 2
  • Gili Solomon
    • 1
  • Zvi Hayouka
    • 1
  • Arieh Gertler
    • 1
    Email author
  1. 1.The Robert H. Smith Faculty of Agriculture, Food and EnvironmentThe Hebrew University of JerusalemRehovotIsrael
  2. 2.Department of Animal Physiology and EndocrinologyThe University of Agriculture in KrakowKrakowPoland

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