title: Purification and characterization of recombinant human interleukin-1 beta produced in Escherichia coli.
abstract: Recombinant human interleukin-1 beta (rIL-1 beta) produced in Escherichia coli was purified to homogeneity by a combination of mass ion exchange column chromatography, ion exchange and gel filtration high performance liquid chromatography. The purified rIL-1 beta had a molecular weight of 18 kD on SDS-polyacrylamide gel electrophoresis and an isoelectric point of 6.9 on analytical isoelectric focusing. These values were almost same as those of natural interleukin-1 beta. The amino acid composition and amino acid sequence of the amino terminal region were consistent with those deduced from the cDNA sequence. In addition, the primary structure was confirmed by peptide mapping with lysyl-endopeptidase on reverse phase HPLC. Besides rIL-1 beta with amino terminal Ala, two molecular species, [Met0] rIL-1 beta and [desAla1] rIL-1 beta, were also obtained. Biological and physicochemical properties of the three species of rIL-1 beta were compared.
authors.forename: Y
authors.initials: Y
authors.lastname: Kikumoto
authors.forename: Y M
authors.initials: YM
authors.lastname: Hong
authors.forename: T
authors.initials: T
authors.lastname: Nishida
authors.forename: S
authors.initials: S
authors.lastname: Nakai
authors.forename: Y
authors.initials: Y
authors.lastname: Masui
authors.forename: Y
authors.initials: Y
authors.lastname: Hirai
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001681:Biological Assay; D003001:Cloning, Molecular; D004926:Escherichia coli; D005786:Gene Expression Regulation; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D007526:Isoelectric Point; D008970:Molecular Weight; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000302:isolation & purification*; D013329:Structure-Activity Relationship
chemical_list: D000596:Amino Acids; D007375:Interleukin-1; D011994:Recombinant Proteins