Purification of cytosolic cAMP-independent protein kinases from rat ventral prostate.
Purification of cytosolic cAMP-independent protein kinases from rat ventral prostate.
复制标题
从大鼠腹侧前列腺中纯化胞质 cAMP 独立蛋白激酶。
DOI：
10.1016/0020-711x(86)90068-6
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Ahmed,K
中科院分区：
文献类型：
--
作者：
Goueli,SA;Ferkul,KM;Ahmed,K
从大鼠腹侧前列腺和肝胞质溶胶中纯化了两种cAMP非依赖性蛋白激酶，并将其命名为PK-C1和PK-C2，以将其与前一篇论文中描述的核蛋白激酶区分开来。每种前列腺酶的产率约为5%，每种肝酶的产率约为10%。蛋白激酶C1和C2的前列腺酶的平均纯化倍数分别为1892和3176。它们对酪蛋白的平均比活性分别为40，111和67，340 nmol 32 P掺入/hr/mg酶蛋白。蛋白激酶C1包含一个Mr为39，000的多肽，其在Mg 2 ++ ATP存在下进行磷酸化。蛋白激酶C2由三种多肽组成，分子量分别为41，000、38，000和26，000。其中只有Mr 26，000多肽是自磷酸化的。蛋白激酶C1和C2的Mg 2+需求量为1至4 mM，这取决于蛋白底物的性质。这两种酶都受到100-200 mM NaCl的刺激。Km为ATP的C1和C2激酶为0.01 mM; GTP可以使用的蛋白激酶C2，但具有显着较低的亲和力。酶对酪蛋白，脱磷卵黄蛋白，脱磷卵黄蛋白，精胺结合蛋白在体外的活性，但表现出对组蛋白的活性很小。尽管胞质蛋白激酶C1和C2与核蛋白激酶N1和N2的这些一般性质有一些相似之处，但也注意到一些差异。
Two cAMP-independent protein kinases were purified from rat ventral-prostate and liver cytosol, and were designated PK-C1 and PK-C2 to distinguish them from the nuclear protein kinases described in the preceding paper. The yield of the prostate enzymes was about 5% each, and about 10% each for the liver enzymes. The average fold purification of the prostatic enzymes was 1892 and 3176 for protein kinase C1 and C2, respectively. Their average respective specific activity towards casein was 40,111 and 67,340 nmol 32P incorporated/hr per mg of enzyme protein. protein kinase C1 comprised one polypeptide of Mr 39,000 which underwent phosphorylation in the presence of Mg2++ ATP. Protein kinase C2 comprised three polypeptides of Mr 41,000; 38,000; 26,000. Of these only the Mr 26,000 polypeptide was autophosphorylated. The Mg2+ requirement for protein kinase C1 and C2 was between 1 and 4 mM depending on the nature of the protein substrate. Both enzymes were stimulated by 100-200 mM NaCl. Km for ATP for C1 and C2 kinases was 0.01 mM; GTP could be used only by protein kinase C2 but with a markedly lower affinity. The enzymes were active towards casein, phosvitin, dephosphophosvitin, and spermine-binding protein in vitro, but demonstrated little activity towards histones. Despite several similarities in these general properties of cytosolic protein kinases C1 and C2 with those of nuclear protein kinases N1 and N2, a number of differences are also noted.