Sola M M, Langan T, Cohen P
Department of Biochemistry, University of Dundee, U.K.
Biochim Biophys Acta. 1991 Sep 3;1094(2):211-6. doi: 10.1016/0167-4889(91)90011-l.
The protein phosphatase activity in rat liver cytosol or nuclear extracts that dephosphorylates histone H1 which has been phosphorylated by p34cdc2 is inhibited completely by okadaic acid, but unaffected by inhibitor-2 or magnesium ions, demonstrating that the only enzyme in this tissue capable of dephosphorylating this substrate is a type 2A phosphatase. Fractionation of the cytosol by anion-exchange chromatography and gel filtration demonstrated that histone H1 phosphatase activity coeluted with the major species of protein phosphatase 2A, termed PP2A1 and PP2A2. PP2A1 was the most active histone H1 phosphatase, its histone phosphatase phosphorylase phosphatase activity ratio being 6-fold higher than PP2A2 and 30-fold higher than the free catalytic subunit PP2AC. It is concluded that PP2A1 is likely to be the enzyme which dephosphorylates p34cdc2-labelled histone H1 in vivo and that the A and B subunits which interact with PP2AC in this species each play a key role in facilitating dephosphorylation of this substrate. The results demonstrate that PP2A, in addition to being involved in suppressing the activation of p34cdc2 in vivo, can also function to reverse at least one of its actions.
大鼠肝脏胞质溶胶或核提取物中能使已被p34cdc2磷酸化的组蛋白H1去磷酸化的蛋白质磷酸酶活性被冈田酸完全抑制,但不受抑制剂-2或镁离子的影响,这表明该组织中唯一能够使该底物去磷酸化的酶是2A型磷酸酶。通过阴离子交换色谱和凝胶过滤对胞质溶胶进行分级分离表明,组蛋白H1磷酸酶活性与主要的蛋白质磷酸酶2A(称为PP2A1和PP2A2)共洗脱。PP2A1是活性最高的组蛋白H1磷酸酶,其组蛋白磷酸酶/磷酸化酶磷酸酶活性比分别比PP2A2高6倍,比游离催化亚基PP2AC高30倍。得出的结论是,PP2A1可能是体内使p34cdc2标记的组蛋白H1去磷酸化的酶,并且在该物种中与PP2AC相互作用的A和B亚基在促进该底物的去磷酸化过程中均发挥关键作用。结果表明,PP2A除了参与体内抑制p34cdc2的激活外,还可以发挥作用来逆转其至少一种作用。