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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
10
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pubmed:dateCreated |
2000-10-30
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pubmed:abstractText |
Interleukin (IL-)18 is an activator of NK cells and a co-inducer of Th(1)cytokines, sharing structural features with the IL-1 family of proteins. Unlike most other cytokines, IL-18 and IL-1beta lack a signal peptide, have an all beta-pleated sheet structure and are synthesized as biologically inactive precursors (pro-IL-18 and pro-IL-1beta). These precursors are cleaved by caspase-1 (IL-1beta-converting enzyme, ICE) to form the biologically active mature cytokines. Direct expression of mature recombinant human IL-18 in E. coli resulted in a partially active cytokine. We tested the possibility that correct folding of huIL-18 requires its prior synthesis as pro-IL-18. Because caspase-1 is not readily available, we constructed an expression vector encoding human pro-IL-18 in which the caspase-1 cleavage site was mutated into a factor Xa site. To facilitate purification, the mutated pro-IL-18 cDNA was fused in frame to a glutathione-S-transferase (GST) coding sequence. The GST-pro-IL-18 fusion protein was expressed in E. coli, captured on glutathione agarose and mature human IL-18, exhibiting high biological activity was released upon cleavage with factor Xa. This result indicates that correct folding of huIL-18 occurs at the level of pro-IL-18 and provides a practical way to produce biologically active huIL-18.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
1043-4666
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pubmed:author |
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pubmed:copyrightInfo |
Copyright 2000 Academic Press.
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pubmed:issnType |
Print
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pubmed:volume |
12
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1519-25
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:11023667-Amino Acid Sequence,
pubmed-meshheading:11023667-Animals,
pubmed-meshheading:11023667-Base Sequence,
pubmed-meshheading:11023667-Biological Assay,
pubmed-meshheading:11023667-Caspase 1,
pubmed-meshheading:11023667-Cell Line,
pubmed-meshheading:11023667-Cloning, Molecular,
pubmed-meshheading:11023667-DNA, Complementary,
pubmed-meshheading:11023667-Dose-Response Relationship, Drug,
pubmed-meshheading:11023667-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:11023667-Escherichia coli,
pubmed-meshheading:11023667-Factor Xa,
pubmed-meshheading:11023667-Glutathione,
pubmed-meshheading:11023667-Glutathione Transferase,
pubmed-meshheading:11023667-Humans,
pubmed-meshheading:11023667-Interferon-gamma,
pubmed-meshheading:11023667-Interleukin-18,
pubmed-meshheading:11023667-Leukocytes, Mononuclear,
pubmed-meshheading:11023667-Mice,
pubmed-meshheading:11023667-Models, Genetic,
pubmed-meshheading:11023667-Molecular Sequence Data,
pubmed-meshheading:11023667-Mutation,
pubmed-meshheading:11023667-Plasmids,
pubmed-meshheading:11023667-Protein Folding,
pubmed-meshheading:11023667-Protein Precursors,
pubmed-meshheading:11023667-Recombinant Fusion Proteins,
pubmed-meshheading:11023667-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:11023667-Time Factors
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pubmed:year |
2000
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pubmed:articleTitle |
Production of a biologically active human interleukin 18 requires its prior synthesis as PRO-IL-18.
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pubmed:affiliation |
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, 76100, Israel.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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