Kikumoto Y, Hong Y M, Nishida T, Nakai S, Masui Y, Hirai Y
Biochem Biophys Res Commun. 1987 Aug 31;147(1):315-21. doi: 10.1016/s0006-291x(87)80123-7.
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.
通过大规模离子交换柱色谱、离子交换和凝胶过滤高效液相色谱相结合的方法，对在大肠杆菌中产生的重组人白细胞介素-1β（rIL-1β）进行纯化，直至达到同质。纯化后的rIL-1β在SDS-聚丙烯酰胺凝胶电泳上的分子量为18 kD，在分析等电聚焦上的等电点为6.9。这些值与天然白细胞介素-1β的几乎相同。氨基末端区域的氨基酸组成和氨基酸序列与从cDNA序列推导的一致。此外，通过在反相HPLC上用赖氨酰内肽酶进行肽图谱分析，证实了一级结构。除了氨基末端为丙氨酸的rIL-1β外，还获得了两种分子形式，即[Met0] rIL-1β和[desAla1] rIL-1β。比较了三种rIL-1β的生物学和物理化学性质。
