Analysis and characterisation of IL-1 beta processing in rainbow trout, Oncorhynchus mykiss

S H Hong, Jun Zou, Bertrand Collet, N C Bols, Christopher John Secombes

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Mammalian IL-1beta is produced as a biologically inactive 31 kDa precursor, which is converted to the active 18 kDa form by proteolytic processing. Synthesis and processing of native piscine IL-1beta is poorly understood. In the present study, the native IL-1beta precursor or mature peptides were detected at sizes of approx. 29 kDa and 24 kDa in cell lysates of a rainbow trout macrophage cell line RTS-11, with or without LPS stimulation, by Western blot analysis using a polyclonal antibody against the putative trout mature IL-1beta (rmIL-1beta) produced in Escherichia coli. Processing of the 29 kDa precursor into a 24 kDa mature peptide was confirmed by analysis of such proteins using a monoclonal conjugate (Ni-NTA-HRP) against 6 histidines in lysates of the RTS-11 cells transfected with an expression plasmid containing the IL-1beta precursor molecule tagged with 6 histidines at its C terminus. Only the recombinant mature 24 kDa) IL-1beta/HIS protein was purified from the culture supernatants of the transfected cells, indicating the molecule is cleaved to be secreted. These findings strongly suggest that the trout IL-1beta molecule is processed in trout macrophages in an analogous way to the situation with mammalian IL-1beta despite the lack of clear ICE cut site. (C) 2003 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)453-459
Number of pages7
JournalFish & Shellfish Immunology
Volume16
Issue number3
Early online date28 Sep 2003
DOIs
Publication statusPublished - Mar 2004

Keywords

  • interleukin-1 beta
  • processing
  • ICE-independent
  • rainbow trout
  • converting-enzyme
  • cysteine protease
  • cell-line
  • IL-1 beta
  • interleukin-1-beta
  • precursor
  • monocytes
  • cleavage
  • produce

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