PROTEIN KINASE INDUCTION IN ESCHERICHIA-COLI BY BACTERIOPHAGE T7
PROTEIN KINASE INDUCTION IN ESCHERICHIA-COLI BY BACTERIOPHAGE T7
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DOI：
10.1073/pnas.71.2.586
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发表时间：
1974-01-01
影响因子：
11.1
通讯作者：
STUDIER, FW
中科院分区：
文献类型：
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作者：
RAHMSDORF, HJ;PAI, SH;STUDIER, FW
在T7噬菌体感染后，可以显示蛋白激酶(EC 2.7.1.37；ATP：蛋白磷酸转移酶)的活性。活体十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和放射自显影。无细胞提取物催化[γ-32P]三磷酸腺苷的末端磷酸基转移到内源性蛋白受体或添加的组蛋白。磷酸与蛋白质之间的结合具有磷酸丝氨酸的特征：在1N HCl(100°)中稳定，并被1N KOH(37°)和碱性磷酸酶处理所切割。此外，在部分酸解后，放射性磷酸盐在聚乙烯亚胺-纤维素薄层色谱图上与标记O-磷酸丝氨酸一起迁移。未感染细胞中的酶活性可以忽略不计。紫外线照射噬菌体基因组可以阻止蛋白激酶的出现，而照射宿主基因组则不能。该酶的活性在感染后4分钟出现，其基因图谱位于基因启动子1附近的早期区域。核糖体蛋白在体内被磷酸化，在体外是底物。添加环AMP或环GMP不改变体外酶的活性。
After bacteriophage T7 infection, a protein kinase (EC 2.7.1.37; ATP:protein phosphotransferase) activity can be demonstrated inE. coli in vivoby sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Cell-free extracts catalyzed the transfer of the terminal phosphoryl group of [γ-32P]ATP to endogenous protein acceptor or to added histone. The bond between phosphate and protein shows the characteristics of serine phosphate: it is stable in 1 N HCl (100°) and cleaved by 1 N KOH (37°) and by alkaline phosphatase treatment. Moreover, after partial acid hydrolysis, radiophosphate migrates with markerO-phosphoserine on polyethyleneimine-cellulose thin-layer chromatograms. Enzyme activity in uninfected cells is negligible. Ultraviolet irradiation of the phage genome prevents the appearance of the protein kinase; irradiation of the host genome does not. The enzyme activity occurs 4 min after infection and its gene maps in the early region (promoter proximal to gene1). Ribosomal proteins are phosphorylatedin vivoand are substratesin vitro. Enzyme activityin vitrois not changed by addition of cyclic AMP or cyclic GMP.