Purification and biochemical characterization of an invertebrate interleukin 1.
Purification and biochemical characterization of an invertebrate interleukin 1.
复制标题
无脊椎动物白介素 1 的纯化和生化表征。
DOI：
10.1016/0161-5890(91)90126-5
复制
发表时间：
1991
影响因子：
3.6
通讯作者：
Habicht,GS
中科院分区：
文献类型：
--
作者：
Beck,G;Habicht,GS
白细胞介素1 （IL-1）是巨噬细胞释放的一种主要免疫调节蛋白，具有许多与宿主防御相关的特性。IL-1在无脊椎动物中被发现，证明了它在体内平衡中的重要性。将脊椎动物蛋白与无脊椎动物蛋白相比较的第一步是纯化待研究的蛋白质。从海星体腔液中分离得到IL-1，并对其进行了均质纯化。IL-1的等电点分别为7.4、5.4和4.8。pI 4.8种的分子量为22000，pI 7.4和pI 5.4种的分子量均为17000。较高的mrform也被发现。这些分子在人类黑色素瘤A375细胞毒性IL-1试验中具有生物活性，并且能够刺激小鼠真皮成纤维细胞增殖、蛋白质合成和pge2的产生。对pI 4.8和pI 5.4进行了纯化，并测定了氨基酸组成。pI 4.8和pI 5.4的比活性分别为3 × 106和1 × 106单位mg−1，达到200倍以上。分离出pI 7.4型，进行部分n端序列分析。动物和无脊椎动物IL-1在分子量、等电点和生物学特性上的相似性表明它是一种重要的、进化稳定的宿主防御分子。
Interleukin 1 (IL-1) is a major immunoregulatory protein released by macrophages with many host defense related properties. That IL-1 has been found in the invertebrates attests to its importance in homeostasis.The first step in comparing the vertebrate protein to its invertebrate correlate is to purify the protein to study. We have purified to homogeneity IL-1 isolated from the coelomic fluid of the starfishAsterias forbesi. The IL-1 had isoelectric points of 7.4, 5.4 and 4.8. The pI 4.8 species had a molecular weight of 22,000 and the pI 7.4 and 5.4 species both had Mrof 17,000. Higher Mrforms were also found. These molecules were biologically active in the human melanoma A375 cytotoxity assay for IL-1, and were also able to stimulate murine dermal fibroblast proliferation, protein synthesis, and PGE2production. The pI 4.8 and 5.4 forms were purified to homogeneity and the amino acid composition was determined. The pI 4.8 and 5.4 species were purified more than 200-fold to specific activities of 3 × 106and 1 × 106units mg−1, respectively. The pI 7.4 form was isolated and partialN-terminal sequence analysis was performed. The similarities of molecular weight, isoelectric points and biological properties between verebrate and invertebrate IL-1 show that it is an important, evolutionarily stable host defense molecule.