Purification and partial sequence of human osteoclast-activating factor: identity with interleukin 1 beta.
Purification and partial sequence of human osteoclast-activating factor: identity with interleukin 1 beta.
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DOI：
10.4049/jimmunol.135.4.2562
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发表时间：
1985-10
影响因子：
4.4
通讯作者：
F. Dewhirst;P. Stashenko;J. Mole;T. Tsurumachi
中科院分区：
文献类型：
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作者：
F. Dewhirst;P. Stashenko;J. Mole;T. Tsurumachi
将淋巴因子破骨细胞活化因子（OAF）纯化至均一。在无血清培养条件下，用伴刀豆球蛋白A和佛波酯肉豆蔻酸酯刺激人外周血单个核细胞产生OAF。OAF通过连续凝胶过滤、离子交换和反相HPLC纯化，通过以下骨吸收活性。通过银染聚丙烯酰胺凝胶上单个17，800-道尔顿条带、等电聚焦上单个pI 6.8条带和单个氨基末端序列的标准表明均一性。纯化的OAF在约0.66 ng/ml的浓度下刺激胎鼠长骨的钙的半数最大释放。OAF的氨基末端序列被确定，并发现是相同的白细胞介素1 β。在胸腺细胞增殖试验中，均质OAF具有8.2 × 10（6）U/mg的活性。因为海军陆战队，同种OAF和白细胞介素1 β在胸腺细胞增殖试验中的等电点、氨基末端序列和比活性是相同的，我们得出结论，它们是相同的分子，白细胞介素1 β是凝集素刺激的外周血单核细胞产生的具有OAF活性的主要蛋白质。
The lymphokine osteoclast-activating factor (OAF) was purified to homogeneity. OAF was produced by human peripheral blood mononuclear cells stimulated with concanavalin A and phorbol myristate acetate under serum-free culture conditions. OAF was purified by sequential gel filtration, ion-exchange, and reverse-phase HPLC by following bone resorptive activity. Homogeneity was indicated by the criteria of a single 17,800-dalton band on silver-stained polyacrylamide gels, a single pI 6.8 band on isoelectric focusing, and a single aminoterminal sequence. Purified OAF stimulated half-maximal release of calcium from fetal rat long bones at a concentration of approximately 0.66 ng/ml. The amino-terminal sequence of OAF was determined and found to be identical to that of interleukin 1 beta. Homogeneous OAF possessed an activity of 8.2 X 10(6) U/mg in the thymocyte proliferation assay. Because the m.w., isoelectric point, amino-terminal sequence, and specific activity in the thymocyte proliferation assay are the same for homogeneous OAF and interleukin 1 beta, we conclude that they are the same molecule, and that interleukin 1 beta is the major protein with OAF activity produced by lectin-stimulated peripheral blood mononuclear cells.