Comparative biochemical properties of p21 ras molecules coded for by viral and cellular ras genes
Comparative biochemical properties of p21 ras molecules coded for by viral and cellular ras genes
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病毒和细胞 ras 基因编码的 p21 ras 分子的生化特性比较
DOI：
10.1128/jvi.44.2.509-519.1982
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发表时间：
1982
影响因子：
5.4
通讯作者：
E. Scolnick
中科院分区：
文献类型：
--
作者：
A. Papageorge;D. Lowy;E. Scolnick
在早期的研究中，我们分子克隆了一个正常细胞基因c-rasH-1，与哈维鼠肉瘤病毒（v-rasH）的v-ras癌基因同源。通过将 c 型逆转录病毒启动子连接到 c-rasH-1，我们可以用 c-rasH-1 基因转化 NIH 3T3 细胞。转化的细胞含有高水平的由 c-rasH-1 基因编码的 p21 蛋白。在目前的研究中，我们广泛纯化了 v-rasH p21 和 c-rasH p21，并比较了这两种 p21 分子的体内和体外生化特性。 v-rasH 和 c-rasH-1 编码的 p21 蛋白具有某些共同特性：每种蛋白均作为前体蛋白合成，随后与质膜的内表面结合；每种蛋白质都与鸟嘌呤核苷酸结合活性相关，这种特性可在高压液相色谱分子筛分柱上与 p21 分子共纯化。在一些其他特性中，v-rasH 和 c-rasH p21 蛋白有所不同。在体内，大约 20% 至 30% 的 v-rasH p21 分子是以含有磷酸苏氨酸的 pp21 分子的形式存在的，而在体内，只有一小部分 c-rasH-1 p21 含有磷酸盐，并且这种磷酸盐是在丝氨酸残基上发现的。具有真实磷酸苏氨酸肽的 v-rasH pp21 分子可以通过自磷酸化反应在体外合成，其中 GTP 的 γ 磷酸盐被转移到 v-rasH p21。体外纯化的 c-rasH-1 p21 没有自磷酸化活性。结果表明 v-rasH 和 c-rasH-1 编码的 p21 蛋白之间存在重大的质量差异。由小鼠源性致癌病毒 BALB 鼠肉瘤病毒编码的 p21 与 c-rasH-1 编码的 p21 相似，因为它结合鸟嘌呤核苷酸，但没有明显用 32Pi 标记。对ras基因家族其他成员编码的p21的形式进行了比较，结果表明鸟嘌呤核苷酸结合活性对于ras基因家族所有已知成员编码的p21分子是共有的。
In earlier studies, we molecularly cloned a normal cellular gene, c-rasH-1, homologous to the v-ras oncogene of Harvey murine sarcoma virus (v-rasH). By ligating a type c retroviral promotor to c-rasH-1, we could transform NIH 3T3 cells with the c-rasH-1 gene. The transformed cells contained high levels of a p21 protein coded for by the c-rasH-1 gene. In the current studies, we have purified extensively both v-rasH p21 and c-rasH p21 and compared the in vivo and in vitro biochemical properties of both these p21 molecules. The p21 proteins coded for by v-rasH and c-rasH-1 shared certain properties: each protein was synthesized as a precursor protein which subsequently became bound to the inner surface of the plasma membrane; each protein was associated with guanine nucleotide-binding activity, a property which copurified with p21 molecules on a high-pressure liquid chromatography molecular sizing column. In some other properties, the v-rasH and c-rasH p21 proteins differed. In vivo, approximately 20 to 30% of v-rasH p21 molecules were in the form of phosphothreonine-containing pp21 molecules, whereas in vivo only a minute fraction of c-rasH-1 p21 contained phosphate, and this phosphate was found on a serine residue. v-rasH pp21 molecules with an authentic phosphothreonine peptide could be synthesized in vitro in an autophosphorylation reaction in which the gamma phosphate of GTP was transferred to v-rasH p21. No autophosphorylating activity was associated with purified c-rasH-1 p21 in vitro. The results indicate a major qualitative difference between the p21 proteins coded for by v-rasH and c-rasH-1. The p21 coded for by a mouse-derived oncogenic virus, BALB murine sarcoma virus, resembled the p21 coded for by c-rasH-1 in that it bound guanine nucleotides but did not label appreciably with 32Pi. The forms of p21 coded for by other members of the ras gene family were compared, and the results indicate that the guanine nucleotide-binding activity is common to p21 molecules coded for by all known members of the ras gene family.
通过蛋白激酶级联调节艾氏腹水肿瘤 Na K -ATP 酶 β 亚基的磷酸化。
DOI：
--
发表时间：
1981
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Spector,M;O'Neal,S;Racker,E
通讯作者：
Racker,E