title: Purification and characterization of human interleukin-1 beta expressed in recombinant Escherichia coli.
abstract: The high-level expression of human interleukin-1 beta in Escherichia coli is described. The protein contributes about 12% of the total cell protein and is associated with the soluble cytoplasmic fraction of the cell. A method for the purification of the protein is given which is based on anion- and cation-exchange chromatographies. The isolated protein, shown to be homogeneous by several analytical methods, has been characterized by amino acid analysis, N- and C-terminal sequence analysis and analytical centrifugation. The protein is biologically active as demonstrated by two different in vitro assays, namely, the mononuclear cell factor (IL-1/MCF) assay and lymphocyte activating factor (IL-1/LAF) assay. The specific activities determined with the IL-1/MCF and IL-1/LAF assays, are 2 X 10(7) and 4 X 10(7) units mg-1, respectively.
mesh_terms: D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D015232:Dinoprostone; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D003012:Microbial Collagenase / Q000378:metabolism; D008970:Molecular Weight; D010957:Plasmids; D011458:Prostaglandins E / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D012867:Skin / Q000187:drug effects / Q000378:metabolism
chemical_list: D007375:Interleukin-1; D011458:Prostaglandins E; D011994:Recombinant Proteins; D004247:DNA; D003012:Microbial Collagenase; D015232:Dinoprostone