Molecular and functional characterization of a fish inducible-type nitric oxide synthase

Jeroen Saeij, René J M Stet, Adrie Groeneveld, Lidy B M Verburg-Van Kemenade, Willem B. Van Muiswinkel, Geert F. Wiegertjes

Research output: Contribution to journalArticle

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Abstract

Using an oligonucleotide primer based on a partial goldfish inducible nitric oxide synthase (iNOS) sequence, a complete carp iNOS cDNA was isolated from an activated carp phagocyte cDNA library. Nucleotide and predicted amino acid sequence analysis indicate that carp iNOS encodes a 1127-amino acid protein with 57% sequence identity to human iNOS. Like mammalian NOSs, carp iNOS protein contains putative binding sites for heme, tetrahydrobiopterin, calmodulin, flavine mononucleotide, flavine adenine dinucleotide, and NADPH. Phylogenetic analysis, using neighbor joining, showed that the carp iNOS protein clustered together with the other vertebrate iNOS proteins. Inducibility of carp iNOS was confirmed by reverse transcription-polymerase chain reaction after stimulation of carp phagocytes with lipopolysaccharide or the protozoan blood flagellate Trypanoplasma borreli. These stimulators produced high amounts of nitric oxide that were toxic for T. borreli in vitro. The nuclear transciption factor NF-κB was shown to play a role in the induction of iNOS transcription.

Original languageEnglish (US)
Pages (from-to)339-346
Number of pages8
JournalImmunogenetics
Volume51
Issue number4-5
StatePublished - 2000
Externally publishedYes

Fingerprint

Nitric Oxide Synthase Type II
Carps
Fishes
Phagocytes
Proteins
Goldfish
DNA Primers
Poisons
Protein Sequence Analysis
Adenine
Calmodulin
Heme
NADP
Gene Library
Reverse Transcription
Lipopolysaccharides
Vertebrates
Nitric Oxide
Nucleotides
Complementary DNA

Keywords

  • Evolution
  • Fish
  • iNOS
  • Parasite
  • Transcription

ASJC Scopus subject areas

  • Immunology
  • Genetics

Cite this

Saeij, J., Stet, R. J. M., Groeneveld, A., Verburg-Van Kemenade, L. B. M., Van Muiswinkel, W. B., & Wiegertjes, G. F. (2000). Molecular and functional characterization of a fish inducible-type nitric oxide synthase. Immunogenetics, 51(4-5), 339-346.

Molecular and functional characterization of a fish inducible-type nitric oxide synthase. / Saeij, Jeroen; Stet, René J M; Groeneveld, Adrie; Verburg-Van Kemenade, Lidy B M; Van Muiswinkel, Willem B.; Wiegertjes, Geert F.

In: Immunogenetics, Vol. 51, No. 4-5, 2000, p. 339-346.

Research output: Contribution to journalArticle

Saeij, J, Stet, RJM, Groeneveld, A, Verburg-Van Kemenade, LBM, Van Muiswinkel, WB & Wiegertjes, GF 2000, 'Molecular and functional characterization of a fish inducible-type nitric oxide synthase', Immunogenetics, vol. 51, no. 4-5, pp. 339-346.
Saeij J, Stet RJM, Groeneveld A, Verburg-Van Kemenade LBM, Van Muiswinkel WB, Wiegertjes GF. Molecular and functional characterization of a fish inducible-type nitric oxide synthase. Immunogenetics. 2000;51(4-5):339-346.
Saeij, Jeroen ; Stet, René J M ; Groeneveld, Adrie ; Verburg-Van Kemenade, Lidy B M ; Van Muiswinkel, Willem B. ; Wiegertjes, Geert F. / Molecular and functional characterization of a fish inducible-type nitric oxide synthase. In: Immunogenetics. 2000 ; Vol. 51, No. 4-5. pp. 339-346.
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