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首页> 外文期刊>Molecular and Cellular Biology >Purified polyoma virus medium T antigen has tyrosine-specific protein kinase activity but no significant phosphatidylinositol kinase activity.
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Purified polyoma virus medium T antigen has tyrosine-specific protein kinase activity but no significant phosphatidylinositol kinase activity.

机译：纯化的多瘤病毒培养基T抗原具有酪氨酸特异性蛋白激酶活性，但没有明显的磷脂酰肌醇激酶活性。

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Medium T antigen, the transforming protein of polyoma virus, is associated with pp60c-src and strongly activates its tyrosine-specific protein kinase activity. We investigated whether the medium T-pp60c-src complex is also associated with an activity that phosphorylates the membrane phospholipid phosphatidylinositol, as shown for pp60v-src and p68v-ros, the transforming proteins of Rous sarcoma virus and avian sarcoma virus UR2, respectively. Medium T was purified by affinity chromatography from extracts of polyoma virus-infected mouse fibroblasts. It was bound to antibodies against a peptide corresponding to the carboxy terminus of medium T and released from the immune complex with an excess of the same peptide. In a second step, the partially purified medium T was bound to antibodies against another peptide corresponding to an internal region of medium T and released with excess peptide. Further purification was carried out with a monoclonal antibody against pp60c-src. Samples from each purification step were examined for protein kinase and phosphatidylinositol kinase activity. The highly purified preparations of the medium T-pp60c-src complex showed very low levels of phosphatidylinositol kinase activity, and no difference between medium T from transforming viruses and nontransforming hr-t mutants was detected. In contrast, protein kinase activity was associated with medium T purified from transforming viruses but not from hr-t mutants.
机译：中T抗原是多瘤病毒的转化蛋白，与pp60c-src相关，并强烈激活其酪氨酸特异性蛋白激酶活性。我们研究了培养基T-pp60c-src复合物是否也与使膜磷脂磷脂酰肌醇磷酸化的活性有关，如分别针对劳斯肉瘤病毒和禽肉瘤病毒UR2的pp60v-src和p68v-ros所示。通过亲和层析从多瘤病毒感染的小鼠成纤维细胞提取物中纯化培养基T。它与针对对应于培养基T的羧基末端的肽的抗体结合，并与过量的相同肽一起从免疫复合物中释放。在第二步骤中，将部分纯化的培养基T与针对对应于培养基T的内部区域的另一种肽的抗体结合，并与过量的肽一起释放。用针对pp60c-src的单克隆抗体进行进一步纯化。检查来自每个纯化步骤的样品的蛋白激酶和磷脂酰肌醇激酶活性。 T-pp60c-src培养基复合物的高纯度制剂显示出极低水平的磷脂酰肌醇激酶活性，并且未检测到来自转化病毒的培养基T和非转化hr-t突变体之间的差异。相反，蛋白激酶活性与从转化病毒而非从hr-t突变体纯化得到的培养基T有关。

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