Schmitt M, Kempf J, Quirin-Stricker C
Biochim Biophys Acta. 1977 Apr 12;481(2):438-49. doi: 10.1016/0005-2744(77)90277-7.
Three different types of protein kinases (ATP: protein phosphotransferase, EC 2.7.1.37) were isolated and partially purified from a mouse plasmacytoma microsomal KCl wash fraction, then chromatographed on DEAE cellulose and phosphocellulose. The three protein kinase activities designated by protein kinase I, II and III were characterized with respect to their capacity to utilize [gamma-32P]ATP and [gamma-32P]GTP, to interact with cyclic AMP, stimulation by cyclic AMP, substrate specificity and sedimentation behaviour on glycerol gradient centrifugation. Protein kinase I was found to be cyclic AMP dependent and preferentially phosphorylated histones. Protein kinase II and III were insensitive to cyclic AMP, protein kinase II preferentially phosphorylated histones and the protein(s) of a ribosomal KCl wash fraction eluted from DEAE cellulose between 0.2 and 0.35 M KCl and termed "PPx". Protein kinase III phosphorylated casein and ribosomal proteins to a great extent. Studies with glycerol density gradient centrifugation indicated that protein kinase I sediments as a component of about 4.4 S, protein kinase II of 4.3 S and protein kinase III of 3 S. Chromatography on phosphocellulose of the protein kinases isolated from purified free polysomes showed the same type of protein kinases as those from microsomes. So it appears unlikely that protein kinase I and II were contaminants from the cytosol.
从小鼠浆细胞瘤微粒体氯化钾洗涤组分中分离并部分纯化了三种不同类型的蛋白激酶（ATP：蛋白磷酸转移酶，EC 2.7.1.37），然后在DEAE纤维素和磷酸纤维素上进行色谱分析。根据它们利用[γ-32P]ATP和[γ-32P]GTP的能力、与环磷酸腺苷（cAMP）相互作用的能力、cAMP的刺激作用、底物特异性以及在甘油梯度离心中的沉降行为，对分别命名为蛋白激酶I、II和III的三种蛋白激酶活性进行了表征。发现蛋白激酶I依赖于cAMP，且优先磷酸化组蛋白。蛋白激酶II和III对cAMP不敏感，蛋白激酶II优先磷酸化组蛋白以及从DEAE纤维素上在0.2至0.35M氯化钾之间洗脱的核糖体氯化钾洗涤组分中的一种或多种蛋白，这些蛋白被称为“PPx”。蛋白激酶III在很大程度上磷酸化酪蛋白和核糖体蛋白。甘油密度梯度离心研究表明，蛋白激酶I以约4.4S的组分形式沉降，蛋白激酶II以4.3S的形式沉降，蛋白激酶III以3S的形式沉降。从纯化的游离多核糖体中分离的蛋白激酶在磷酸纤维素上的色谱分析显示出与从微粒体中分离的蛋白激酶相同类型的蛋白激酶。因此，蛋白激酶I和II似乎不太可能是来自胞质溶胶的污染物。
