A Comprehensive Bioinformatic Assessment of Different Signal Peptides for Secretory Expression of Human Growth Hormone in Escherichia Coli: An In Silico Study

Document Type : Original Article (s)

Authors

1 MSc Student, Department of Molecular Cell Biology, School of Applied Science, University of Guilan, Rasht, Iran

2 Associate Professor, Department of Molecular Cell Biology, School of Applied Science, University of Guilan, Rasht, Iran

3 Assistant Professor, Department of Biotechnology, School of Agriculture, University of Guilan, Rasht, Iran

4 Associate Professor, Department of Biotechnology, School of Agriculture, University of Guilan, Rasht, Iran

Abstract

Background: Nowadays, by genetic engineering and bioinformatics, large scale production of pharmacological recombinant proteins in Escherichia coli (E. coli) bacteria, which has unique expression properties, becomes a routine and economic imperative. In this study, periplasmic production of human growth hormone was investigated using bioinformatics methods.Methods: The aim of this study was bioinformatic evaluation of 48 human signal peptides by reliable servers for expression analysis of human growth hormone in Escherichia coli. Accuracy and precision of 48 signal peptides were evaluated via powerful SignalP server. Physicochemical properties of remaining signal peptides were investigated using Genescript and Protparam servers. Solubility of protein, secretory activity of signal peptides after expression, and transmission mechanism of signal peptides were investigated using Solpro, ProtCompB and PRED-TAT, respectively.Findings: Theoretically, proline rich protein HaeIII subfamily 1 (PRH1), C10orf99, and prolactin-releasing hormone (PRLH) signal peptides were predicted as the most proper signal peptides in fusion of human growth hormone protein, respectively.Conclusion: Secretory expression instead of cytoplasmic expression provides benefits. This study results indicated that by examining different signal peptide sequences in fusion with human growth hormone protein, achieving signal peptides with potential and capability for high expression is possible. The accuracy of these results can be verified in future studies and experiments.

Keywords

References

  1. Reh CS, Geffner ME. Somatotropin in the treatment of growth hormone deficiency and Turner syndrome in pediatric patients: A review. Clin Pharmacol 2010; 2: 111-22.
  2. Franklin SL, Geffner ME. Growth hormone: the expansion of available products and indications. Pediatr Clin North Am 2011; 58(5): 1141-65, x.
  3. Kim MJ, Park HS, Seo KH, Yang HJ, Kim SK, Choi JH. Complete solubilization and purification of recombinant human growth hormone produced in Escherichia coli. PLoS One 2013; 8(2): e56168.
  4. Yoon SH, Kim SK, Kim JF. Secretory production of recombinant proteins in Escherichia coli. Recent Pat Biotechnol 2010; 4(1): 23-9.
  5. Thanassi DG, Hultgren SJ. Multiple pathways allow protein secretion across the bacterial outer membrane. Curr Opin Cell Biol 2000; 12(4): 420-30.
  6. Sockolosky JT, Szoka FC. Periplasmic production via the pET expression system of soluble, bioactive human growth hormone. Protein Expr Purif 2013; 87(2): 129-35.
  7. Zamani M, Nezafat N, Negahdaripour M, Dabbagh F, Ghasemi Y. In silico evaluation of different signal peptides for the secretory production of human growth hormone in E. coli. Int J Pept Res Ther 2015; 21(3): 261-8.
  8. Bendtsen JD, Nielsen H, von Heijne G, Brunak S. Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 2004; 340(4): 783-95.
  9. Petersen TN, Brunak S, von Heijne G, Nielsen H. SignalP 4.0: Discriminating signal peptides from transmembrane regions. Nat Methods 2011; 8(10): 785-6.
  10. Emanuelsson O, Brunak S, von Heijne G, Nielsen H. Locating proteins in the cell using TargetP, SignalP and related tools. Nat Protoc 2007; 2(4): 953-71.
  11. Garcia dlT, Carrasco B, Harding SE. SOLPRO: Theory and computer program for the prediction of SOLution PROperties of rigid macromolecules and bioparticles. Eur Biophys J 1997; 25(5-6): 361-72.
  12. Magnan CN, Randall A, Baldi P. SOLpro: Accurate sequence-based prediction of protein solubility. Bioinformatics 2009; 25(17): 2200-7.
  13. Bendtsen JD, Jensen LJ, Blom N, von HG, Brunak S. Feature-based prediction of non-classical and leaderless protein secretion. Protein Eng Des Sel 2004; 17(4): 349-56.
  14. Bagos PG, Nikolaou EP, Liakopoulos TD, Tsirigos KD. Combined prediction of Tat and Sec signal peptides with hidden Markov models. Bioinformatics 2010; 26(22): 2811-7.
  15. Baneyx F, Mujacic M. Recombinant protein folding and misfolding in Escherichia coli. Nat Biotechnol 2004; 22(11): 1399-408.
  16. Baradaran A, Sieo CC, Foo HL, Illias RM, Yusoff K, Rahim RA. Cloning and in silico characterization of two signal peptides from Pediococcus pentosaceus and their function for the secretion of heterologous protein in Lactococcus lactis. Biotechnol Lett 2013; 35(2): 233-8.
  17. Choi JH, Lee SY. Secretory and extracellular production of recombinant proteins using Escherichia coli. Appl Microbiol Biotechnol 2004; 64(5): 625-35.
  18. Xiong E, Zheng C, Wu X, Wang W. Protein subcellular location: The gap between prediction and experimentation. Plant Mol Biol Rep 2016; 34(1): 52-61.
  19. Mirhoseini Z, Pezeshkian Z, Ghovvati S. Phylogenetic and in silico analysis of interferon beta-1b protein. J Mazandaran Univ Med Sci 2017; 26(145): 70-82. [In Persian].
  20. Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982; 157(1): 105-32.
  21. Chen H, Kim J, Kendall DA. Competition between functional signal peptides demonstrates variation in affinity for the secretion pathway. J Bacteriol 1996; 178(23): 6658-64.
  22. Ikai A. Thermostability and aliphatic index of globular proteins. J Biochem 1980; 88(6): 1895-8.
  23. Guruprasad K, Reddy BV, Pandit MW. Correlation between stability of a protein and its dipeptide composition: A novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Eng 1990; 4(2): 155-61.
  24. Hinsley AP, Stanley NR, Palmer T, Berks BC. A naturally occurring bacterial Tat signal peptide lacking one of the 'invariant' arginine residues of the consensus targeting motif. FEBS Lett 2001; 497(1): 45-9.
Journal of Isfahan Medical School
Volume 35, Issue 440 - Serial Number 440
May and June 2017
Pages 890-899

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History

  • Receive Date: 07 June 2017
  • Revise Date: 25 April 2024
  • Accept Date: 06 April 2022

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