Molecular cloning and functional expression of equine interleukin-1 receptor antagonist.
Equine interleukin-1 receptor antagonist (IL-1ra) was molecularly cloned to establish a basis for cytokine therapy of acute and chronic inflammatory diseases in the horse. cDNA clones encoding the whole coding sequence of equine IL-1ra were isolated from equine peripheral blood mononuclear cells (PBMC) that had been stimulated with lipopolysaccharide (LPS). The equine IL-1ra cDNA obtained in this study contained an open reading frame encoding 177 amino acid residues. The predicted amino acid sequence of equine IL-1ra shared 75.7, 75.3 and 76.3% similarity with sequences of human, murine and rabbit IL-1ras, respectively. An N-glycosylation site and five cysteine residues conserved in human, murine and rabbit IL-1ras were also found at the corresponding positions in equine IL-1ra. Recombinant glutathione S-transferase (GST)-equine IL-1ra fusion protein produced by Escherichia coli was purified. This protein was shown to inhibit the cytostatic or cytotoxic activity of IL-1 on A375S2 cells, indicating that the equine IL-1ra cDNA obtained in this study encodes biologically active equine IL-1ra.
Kato
H
H
Department of Veterinary Internal Medicine, Faculty of Agriculture, University of Tokyo, Japan.
Ohashi
T
T
Matsushiro
H
H
Watari
T
T
Goitsuka
R
R
Tsujimoto
H
H
Hasegawa
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006734:Horse Diseases / Q000628:therapy; D006736:Horses / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007249:Inflammation / Q000628:therapy / Q000662:veterinary; D053590:Interleukin 1 Receptor Antagonist Protein; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000235:genetics / Q000627:therapeutic use; D017386:Sequence Homology, Amino Acid; D012795:Sialoglycoproteins / Q000235:genetics* / Q000627:therapeutic use; D013045:Species Specificity
D017931:DNA Primers; D018076:DNA, Complementary; C507763:IL1RN protein, human; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; D012795:Sialoglycoproteins