Purification and characterization of recombinant human interleukin-1 beta produced in Escherichia coli
Kikumoto, Y.; Hong, Y.M.; Nishida, T.; Nakai, S.; Masui, Y.; Hirai, Y.
Biochemical and Biophysical Research Communications 147(1): 315-321
1987
ISSN/ISBN: 0006-291X PMID: 3307775 DOI: 10.1016/s0006-291x(87)80123-7
Accession: 041135461
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.