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酪氨酸蛋白激酶细胞靶点的鉴定与表征

Identification and characterization of cellular targets for tyrosine protein kinases.

作者信息

Cooper J A, Hunter T

出版信息

J Biol Chem. 1983 Jan 25;258(2):1108-15.

PMID:6571834
Abstract

Protein kinases associated with the transforming proteins of a number of retroviruses are specific for tyrosine. Several proteins in cells transformed by these viruses are phosphorylated at tyrosine. We have now identified three unrelated abundant nonphosphorylated cellular proteins of 46,000, 39,000 and 28,000 daltons in chick embryo cells, which are the unphosphorylated forms of phosphotyrosine-containing proteins and thus are substrates for tyrosine protein kinases. By two-dimensional gel analysis, we have found that the 46,000-dalton protein exists in two unphosphorylated forms of which the more acidic is a minor species. This latter form is phosphorylated, chiefly at serine, in both normal and transformed cells, generating a yet more acidic species. In transformed but not normal cells, the major form is phosphorylated at tyrosine and serine, yielding a fourth isoelectric variant. The 46,000-dalton unphosphorylated protein has been partially purified, and antiserum to it recognizes all four isoelectric variants of the protein. The 39,000-dalton protein has two unphosphorylated forms of which the more acidic is a minor species. The major form is phosphorylated at tyrosine and serine in transformed cells only. The 39,000-dalton unphosphorylated protein has been partially purified, and antiserum raised to it recognizes all three isoelectric variants. The 28,000-dalton protein has a single phosphorylated form which contains serine in normal cells, but both serine and tyrosine in transformed cells.

摘要

与许多逆转录病毒的转化蛋白相关的蛋白激酶对酪氨酸具有特异性。被这些病毒转化的细胞中的几种蛋白质在酪氨酸处被磷酸化。我们现已在鸡胚细胞中鉴定出三种不相关的、分子量分别为46000、39000和28000道尔顿的丰富的非磷酸化细胞蛋白,它们是含磷酸酪氨酸蛋白的未磷酸化形式,因此是酪氨酸蛋白激酶的底物。通过二维凝胶分析,我们发现46000道尔顿的蛋白以两种未磷酸化形式存在,其中酸性较强的是次要形式。后一种形式在正常细胞和转化细胞中主要在丝氨酸处被磷酸化,产生一种酸性更强的形式。在转化细胞而非正常细胞中,主要形式在酪氨酸和丝氨酸处被磷酸化,产生第四种等电变体。46000道尔顿的未磷酸化蛋白已被部分纯化,针对它的抗血清可识别该蛋白的所有四种等电变体。39000道尔顿的蛋白有两种未磷酸化形式,其中酸性较强的是次要形式。主要形式仅在转化细胞中在酪氨酸和丝氨酸处被磷酸化。39000道尔顿的未磷酸化蛋白已被部分纯化,针对它产生的抗血清可识别所有三种等电变体。28000道尔顿的蛋白有一种单一的磷酸化形式,在正常细胞中含有丝氨酸,但在转化细胞中含有丝氨酸和酪氨酸。

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