PRODUCTION, PURIFICATION AND CHARACTERIZATION OF A RABBIT RECOMBINANT IL-1 RECEPTOR ANTAGONIST

被引：21

作者：

MATSUKAWA, A ^{[1
]}

MORI, S ^{[1
]}

OHKAWARA, S ^{[1
]}

MAEDA, T ^{[1
]}

TANASE, S ^{[1
]}

EDAMITSU, S ^{[1
]}

YANAGI, F ^{[1
]}

YOSHINAGA, M ^{[1
]}

机构：

[1] KUMAMOTO UNIV, SCH MED, DEPT BIOCHEM, KUMAMOTO 860, JAPAN

来源：

BIOMEDICAL RESEARCH-TOKYO | 1992年 / 13卷 / 04期

关键词：

D O I：

10.2220/biomedres.13.269

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

1001 ;

摘要：

We describe here the mature translation product of the cDNA for rabbit IL-1 receptor antagonist (IL-1ra) expressed in E.coli. The rabbit recombinant IL-1ra (rrIL-1ra) was purified using DEAE-cellulose and Sephadex G-75, in a conventional chromatography system, followed by a linear NaCl gradient-elution of DEAE-cellulose in an FPLC system. The purified rrIL-1ra was composed of 143 amino acid residues. It had a molecular mass of 19 kD on SDS-PAGE and an isoelectric point of 5.85 on IEF-PAGE. Amino acid sequence analysis of rrIL-1ra showed that the native N-terminal Met was maintained. Specific biological activity of rrIL-1ra was 2.5 x 10(5) units/mg protein, a value indistinguishable from that of the native factor of rabbit inflammatory exudate cells when assessed by using thymocyte co-mitogenic assay. These observations suggest that rrIL-1ra emulates properties of the native protein. This rrIL-1ra manifested inhibition of rabbit rIL-1-beta- and human rIL-1-alpha- and rIL-1-beta-induced proliferation of mouse thymocytes and also inhibited IL-1 activities in culture supernatants of mononuclear cells from animals, including the rabbit, rat, mouse, guinea pig and from human. Thus, rrIL-1ra provides a useful tool for testing IL-1 activities exhibited by both IL-1-alpha and beta in a wide range of species of experimental animals.

引用

页码：269 / 277

页数：9

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