Biological characterisation of a recombinant Atlantic salmon type I interferon synthesized in Escherichia coli

Ei Lin Ooi, Noel Verjan, Ikuo Hirono, Tomonori Nochi, Hidehiro Kondo, Takashi Aoki, Hiroshi Kiyono, Yoshikazu Yuki

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

Abstract

Type I (α/β) interferons (IFNs) are a family of cytokines that stimulate the expression of numerous proteins that mediate an antiviral state in uninfected cells. Two Atlantic salmon (Salmo salar) IFN-α (SasaIFN-α1 & 2) genes have previously been cloned and both were found to contain a putative N-linked glycosylation site. Recombinant SasaIFN-α1 (rSasaIFN-α1) produced in eukaryotic systems has repeatedly been shown to confer antiviral properties. However, different IFN-α subtypes may exhibit differential antiviral activities and be subject to glycosylation. To evaluate the potential therapeutic impact of a rSasaIFN-α, the mature form of the SasaIFN-α2 protein was produced in a high-level Escherichia coli expression system. Expression of the rSasaIFN-α2 was detected by SDS-PAGE and Western blot, and its identity was confirmed by mass spectrometry. In the homologous Chinook salmon embryonic (CHSE-214) cell line, the rSasaIFN-α2 incited early expression of the IFN-induced Mx protein and exhibited high antiviral activity of about 2.8 × 106 U mg-1 against infectious pancreatic necrosis virus (IPNV). Conversely, antiviral protection by rSasaIFN-α2 was not observed in a heterologous Japanese flounder embryo (HINAE) cell line. Hence, a biologically active form of rSasaIFN-α2 was successfully produced using a glycosylation-deficient prokaryotic system and purified to homogeneity, suggesting that glycosylation is not required for its antiviral activity.

Original languageEnglish
Pages (from-to)506-513
Number of pages8
JournalFish and Shellfish Immunology
Volume24
Issue number5
DOIs
Publication statusPublished - 2008 May
Externally publishedYes

Keywords

  • Antiviral activity
  • Atlantic salmon (Salmo salar)
  • Interferon-α
  • Mx protein
  • N-linked glycosylation
  • Recombinant protein

ASJC Scopus subject areas

  • Environmental Chemistry
  • Aquatic Science

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