PURIFICATION AND PROPERTIES OF EXTRACELLULAR SIGNAL-REGULATED KINASE-1, AN INSULIN-STIMULATED MICROTUBULE-ASSOCIATED PROTEIN-2 KINASE
PURIFICATION AND PROPERTIES OF EXTRACELLULAR SIGNAL-REGULATED KINASE-1, AN INSULIN-STIMULATED MICROTUBULE-ASSOCIATED PROTEIN-2 KINASE
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DOI：
10.1021/bi00215a038
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发表时间：
1991-01-08
期刊：
影响因子：
2.9
通讯作者：
COBB, MH
中科院分区：
文献类型：
--
作者：
BOULTON, TG;GREGORY, JS;COBB, MH
在大鼠 1 成纤维细胞中，胰岛素对 MAP2 蛋白激酶或核糖体蛋白 S6 激酶的活性几乎没有或没有刺激作用。相反，在过度表达正常人胰岛素受体的大鼠 1 细胞中（大鼠 1 HIRc B；McClain 等人 (1987) J. Biol. Chem. 262, 14663-14671），胰岛素将 MAP2 和 S6 激酶活性激活接近 5 倍。 通过 Q-Sepharose、苯基-Sepharose、S-Sepharose、磷酸纤维素、QAE-Sepharose、UltrogelAcA54、DEAE-纤维素和第二个 Q-Sepharose 色谱法，从胰岛素处理的大鼠 1 HIRc B 细胞中纯化了 6300 倍以上的 MAP2 激酶。 与 MAP2 相比，其比活性约为 0.8-1-mu-mol.min-1.mg-1；与髓磷脂碱性蛋白相比，其比活性约为 3-mu-mol.min-1.mg-1。 经 SDS-聚丙烯酰胺凝胶电泳，该酶制剂含有 M(r) = 43 000 的一条主带，并通过磷酸酪氨酸抗体进行免疫印迹。 来自 43-kDa 带的序列导致编码该酶的 cDNA 的分离，我们将其命名为 ERK1（细胞外信号调节激酶）（Boulton 等人 (1990) Science 249, 64-67）。
In rat 1 fibroblasts, insulin has little or no stimulatory effect on the activities of either MAP2 protein kinase or ribosomal protein S6 kinase. In contrast, in rat 1 cells that overexpress the normal human insulin receptor (rat 1 HIRc B; McClain et al. (1987) J. Biol. Chem. 262, 14663-14671), insulin activates both MAP2 and S6 kinase activities close to 5-fold. A MAP2 kinase has been purified from insulin-treated rat 1 HIRc B cells over 6300-fold by chromatography on Q-Sepharose, phenyl-Sepharose, S-Sepharose, phosphocellulose, QAE-Sepharose, UltrogelAcA54, DEAE-cellulose, and a second Q-Sepharose. Its specific activity is approximately 0.8-1-mu-mol.min-1.mg-1 with MAP2 and 3-mu-mol.min-1.mg-1 with myelin basic protein. The enzyme preparation contains one major band of M(r) = 43 000 upon SDS-polyacrylamide gel electrophoresis, which is immunoblotted by antibodies to phosphotyrosine. A sequence from the 43-kDa band led to the isolation of a cDNA encoding the enzyme, which we have named ERK1 for extracellular signal-regulated kinase (Boulton et al. (1990) Science 249, 64-67).