Abstract
The gene coding for human interleukin-5 was synthesized and expressed in Escherichia coli under control of a heat-inducible promoter. High-level expression, 10-15% of total cellular protein, was achieved in E. coli. The protein was produced in an insoluble state. A simple extraction, renaturation and purification scheme is described. The recombinant protein was found to be a homodimer, similar to the natural murine-derived protein. Despite the lack of glycosylation, high specific activities were obtained in three 'in vitro' biological assays. Physical characterization of the protein showed it to be mostly alpha-helical, supporting the hypothesis that a conformational similarity exists among certain cytokines.
MeSH terms
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Amino Acids / analysis
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Animals
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Cell Differentiation
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Chemical Phenomena
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Chemistry, Physical
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Disulfides / analysis
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Electrochemistry
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Eosinophils / cytology
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Escherichia coli / metabolism*
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Gene Expression*
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Hot Temperature
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Humans
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Infant, Newborn
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Interleukin-5 / genetics*
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Interleukin-5 / isolation & purification
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Interleukin-5 / pharmacology
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Macromolecular Substances
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Mice
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Molecular Weight
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Promoter Regions, Genetic / genetics
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Protein Conformation
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / pharmacology
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Solubility
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Sulfhydryl Compounds / analysis
Substances
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Amino Acids
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Disulfides
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Interleukin-5
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Macromolecular Substances
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Recombinant Proteins
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Sulfhydryl Compounds