Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs.
cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid sequence of equine IL-1RII demonstrated 70, 60 and 58% similarity with human, mouse and rat sequences, respectively. Recombinant equine soluble IL-1RI and IL-1RII produced in insect cells bound recombinant equine IL-1alpha and IL-1beta. Furthermore, both receptors suppressed the growth inhibitory activities of equine IL-1alpha and IL-1beta toward A375 cells in a dose-dependent manner, indicating that the present equine IL-1RI and IL-1RII cDNA encodes biologically active proteins.
Kirisawa
Rikio
R
Department of Veterinary Microbiology, School of Veterinary Medicine, Rakuno Gakuen University, Bunkyoudai-Midorimachi 582, Ebetsu, Hokkaido 069-8501, Japan. r-kirisa@rakuno.ac.jp
Hashimoto
Norikazu
N
Tazaki
Mizuho
M
Yamanaka
Hitoki
H
Ishii
Risako
R
Hagiwara
Katsuro
K
Iwai
Hiroshi
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D003001:Cloning, Molecular; D006736:Horses / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007375:Interleukin-1 / Q000276:immunology; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D053573:Receptors, Interleukin-1 Type I; D053574:Receptors, Interleukin-1 Type II; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D053573:Receptors, Interleukin-1 Type I; D053574:Receptors, Interleukin-1 Type II; D011994:Recombinant Proteins; D012313:RNA