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病毒转化细胞和未转化细胞中的磷脂酰肌醇激酶活性。

Phosphatidylinositol kinase activity in virus-transformed and nontransformed cells.

作者信息

Sugano S, Hanafusa H

出版信息

Mol Cell Biol. 1985 Sep;5(9):2399-404. doi: 10.1128/mcb.5.9.2399-2404.1985.

DOI:10.1128/mcb.5.9.2399-2404.1985
PMID:2426580
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC366967/
Abstract

We assayed phosphatidylinositol (PI) kinase (EC 2.7.1.67) activity in detergent extracts of nontransformed or virus-transformed cells. Nontransformed chicken embryo fibroblasts (CEF) contain PI kinase activity with an apparent specific activity of 20 pmol/min per mg of protein. This activity sedimented as a single peak with a molecular weight of approximately 60,000 in a glycerol gradient, although immunoprecipitation with anti-p60src sera showed that the PI kinase activity is distinct from p60c-src. Extracts from CEF transformed by Rous sarcoma virus, Fujinami sarcoma virus, or avian sarcoma virus UR2 showed no elevation of PI kinase activity over nontransformed CEF. Removal of the oncogene products from extracts by immunoprecipitation did not change the level of PI kinase activity in extracts, suggesting that putative virus-coded PI kinases do not make a significant contribution to overall levels of PI kinase activity in transformed cells. Additionally, P140gag-fps was separated from cellular PI kinase by phosphocellulose chromatography. This partially purified fraction contained low PI kinase activity distinct from P140gag-fps, indicating that P140gag-fps has no detectable PI kinase activity.

摘要

我们检测了未转化或病毒转化细胞的去污剂提取物中的磷脂酰肌醇（PI）激酶（EC 2.7.1.67）活性。未转化的鸡胚成纤维细胞（CEF）含有PI激酶活性，其表观比活性为每毫克蛋白质20 pmol/分钟。在甘油梯度中，该活性以单个峰的形式沉淀，分子量约为60,000，尽管用抗p60src血清进行免疫沉淀表明PI激酶活性与p60c-src不同。劳斯肉瘤病毒、藤浪肉瘤病毒或禽肉瘤病毒UR2转化的CEF提取物的PI激酶活性与未转化的CEF相比没有升高。通过免疫沉淀从提取物中去除癌基因产物不会改变提取物中PI激酶活性的水平，这表明推测的病毒编码PI激酶对转化细胞中PI激酶活性的总体水平没有显著贡献。此外，通过磷酸纤维素色谱法将P140gag-fps与细胞PI激酶分离。这个部分纯化的级分含有与P140gag-fps不同的低PI激酶活性，表明P140gag-fps没有可检测到的PI激酶活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/4160f2a61549/molcellb00105-0252-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/c37c8b2c3171/molcellb00105-0250-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/d8c88086287f/molcellb00105-0251-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/4160f2a61549/molcellb00105-0252-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/c37c8b2c3171/molcellb00105-0250-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/d8c88086287f/molcellb00105-0251-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fb60/366967/4160f2a61549/molcellb00105-0252-a.jpg

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本文引用的文献

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Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor-promoting phorbol esters.肿瘤促进剂佛波酯对钙激活的、磷脂依赖性蛋白激酶的直接激活作用。
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纯化的多瘤病毒中T抗原具有酪氨酸特异性蛋白激酶活性，但无明显的磷脂酰肌醇激酶活性。
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Proto-oncogene c-ros codes for a molecule with structural features common to those of growth factor receptors and displays tissue specific and developmentally regulated expression.原癌基因c-ros编码一种具有与生长因子受体共同结构特征的分子，并表现出组织特异性和发育调控性表达。
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