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劳氏肉瘤病毒转化的鸡成纤维细胞提取物中36K蛋白的体外磷酸化作用

In vitro phosphorylation of the 36K protein in extract from Rous sarcoma virus-transformed chicken fibroblasts.

作者信息

Kobayashi N, Tanaka A, Kaji A

出版信息

J Biol Chem. 1981 Mar 25;256(6):3053-8.

PMID:6259148
Abstract

When cell-free extracts of chickens embryo fibroblasts transformed by Rous sarcoma virus (RSV) were incubated with [gamma-32P]ATP, a protein having a Mr of 36,000 was phosphorylated. Two-dimensional electrophoresis of a mixture of phosphorylated proteins formed in vitro and in vivo showed that they are indistinguishable. The in vitro phosphorylation of the Mr = 36,000 protein was completely inhibited by serum isolated from rabbit bearing tumor formed by RSV. In addition, phosphorylation of the 36K protein does not occur if the extract was made from fibroblasts transformed by RSV tsNY68 and cultured at 42 degrees C or from fibroblasts infected with transformation defective RSV. The cell free-phosphorylation of 36K protein was dependent on Mg2+ ions but not dependent on exogenously added cyclic AMP.

摘要

当用劳氏肉瘤病毒(RSV)转化的鸡胚成纤维细胞的无细胞提取物与[γ-32P]ATP一起温育时,一种分子量为36,000的蛋白质被磷酸化。体外和体内形成的磷酸化蛋白质混合物的双向电泳表明它们无法区分。从携带RSV形成的肿瘤的兔子中分离的血清可完全抑制分子量为36,000的蛋白质的体外磷酸化。此外,如果提取物由经RSV tsNY68转化并在42℃培养的成纤维细胞制备,或者由感染转化缺陷型RSV的成纤维细胞制备,则不会发生36K蛋白的磷酸化。36K蛋白的无细胞磷酸化依赖于Mg2+离子,但不依赖于外源添加的环磷酸腺苷。

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