A partial sequence for nitric oxide synthase from a goldfish (Carassius auratus) macrophage cell line

K. J. Laing, P. S. Grabowski, M. Belosevic, C. J. Secombes*

*Corresponding author for this work

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Expression of inducible nitric oxide synthase (iNOS) mRNA was detected in a recently developed goldfish macrophage cell line by RT-PCR, using degenerate primers designed against conserved nucleotide motifs within the different mammalian isoforms of NOS. Increased expression of iNOS poststimulation with LPS was found, and suggests that it is a functional enzyme in goldfish macrophages, supporting the view that iNOS regulation is pretranslational. The nucleotide sequence translated in one reading frame with no stop codons to produce a partial peptide containing 164 amino acids, with highest homology (85%) to a recently identified rainbow trout iNOS sequence. The peptide translation also gave an insight into the conservation of binding motifs, since two cofactor binding sites were present in the amplified PCR product (FMN and calmodulin). In addition, a 42 aa motif present in the region just upstream of the FMN binding motif of mammalian endothelial and neuronal NOS isoforms was absent in the translation, in agreement with every published sequence for iNOS. Finally, the translation was used to construct an unrooted phylogenetic tree.

Original languageEnglish
Pages (from-to)374-379
Number of pages6
JournalImmunology and Cell Biology
Volume74
Issue number4
DOIs
Publication statusPublished - Dec 1996

Fingerprint

Goldfish
Nitric Oxide Synthase Type II
Nitric Oxide Synthase
Macrophages
Cell Line
Flavin Mononucleotide
Protein Isoforms
Reading Frames
Polymerase Chain Reaction
Peptides
Nucleotide Motifs
Terminator Codon
Oncorhynchus mykiss
Calmodulin
Binding Sites
Amino Acids
Messenger RNA
Enzymes

Keywords

  • Goldfish
  • Macrophages
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

A partial sequence for nitric oxide synthase from a goldfish (Carassius auratus) macrophage cell line. / Laing, K. J.; Grabowski, P. S.; Belosevic, M.; Secombes, C. J.

In: Immunology and Cell Biology, Vol. 74, No. 4, 12.1996, p. 374-379.

Research output: Contribution to journalArticle

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