Antibodies to Xenopus egg S6 kinase II recognize S6 kinase from progesterone- and insulin-stimulated Xenopus oocytes and from proliferating chicken embryo fibroblasts.
Antibodies to Xenopus egg S6 kinase II recognize S6 kinase from progesterone- and insulin-stimulated Xenopus oocytes and from proliferating chicken embryo fibroblasts.
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非洲爪蟾卵 S6 激酶 II 抗体可识别来自孕酮和胰岛素刺激的非洲爪蟾卵母细胞以及增殖的鸡胚成纤维细胞的 S6 激酶。
DOI：
10.1128/mcb.7.9.3147-3155.1987
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发表时间：
1987
影响因子：
5.3
通讯作者：
Maller,JL
中科院分区：
文献类型：
--
作者：
Erikson,E;Stefanovic,D;Blenis,J;Erikson,RL;Maller,JL
核糖体蛋白S6在孕酮或胰岛素诱导的非洲爪蟾卵母细胞成熟过程中高度磷酸化。我们以前已经从非受精卵中纯化了一种Mr 92，000蛋白，它是一种主要的S6激酶。在本文中，我们确认从十二烷基硫酸钠-聚丙烯酰胺凝胶复性的活性，这种蛋白质是S6激酶。这种酶，称为S6激酶II（S6 K II），用于制备多克隆抗血清。与抗血清和纯化的S6 K II形成的免疫复合物能够表达激酶活性，具有与纯化的酶相同的底物特异性，包括S6 K II自身的自磷酸化。抗血清不与S6激酶I反应，S6激酶I是存在于Xenopuseggs中的另一种主要S6激酶，其在色谱上不同于S6 K II。给卵母细胞施用孕酮导致这些细胞提取物中S6激酶活性增加20至25倍。对提取物进行的免疫复合物激酶测定显示，抗S6 K II血清与经孕酮处理的卵母细胞的S6激酶反应。该抗血清还与胰岛素刺激的卵母细胞中活化的S6激酶发生反应。此外，抗S6 K Ⅱ血清可与血清刺激的或劳斯肉瘤病毒转化的鸡胚成纤维细胞中活化的S6激酶反应。这些结果表明，S6 K II或抗原相关的S6激酶在各种细胞类型中受到促有丝分裂刺激的调节。
Ribosomal protein S6 becomes highly phosphorylated during progesterone- or insulin-induced maturation ofXenopus laevisoocytes. We have previously purified anMr92,000 protein as one of the major S6 kinases fromXenopusunfertilized eggs. In this paper we confirm by renaturation of activity from a sodium dodecyl sulfate-polyacrylamide gel that this protein is an S6 kinase. This enzyme, termed S6 kinase II (S6 K II), was used for the preparation of polyclonal antiserum. Immunocomplexes formed with the antiserum and purified S6 K II were able to express kinase activity with the same substrate specificity as that of the purified enzyme, including autophosphorylation of S6 K II itself. The antiserum did not react with S6 kinase I, another major S6 kinase present inXenopuseggs, which is chromatographically distinct from S6 K II. The administration of progesterone to oocytes resulted in a 20- to 25-fold increase in S6 kinase activity in extracts of these cells. Immunocomplex kinase assays done on extracts revealed that anti-S6 K II serum reacted with S6 kinase from progesterone-treated oocytes. This antiserum also reacted with the activated S6 kinase from insulin-stimulated oocytes. In addition, anti-S6 K II serum reacted with activated S6 kinase from chicken embryo fibroblasts stimulated with serum or transformed by Rous sarcoma virus. These results indicate that S6 K II or an antigenically related S6 kinase(s) is subject to regulation by mitogenic stimuli in various cell types.