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一种也存在于被猿猴病毒40转化的细胞中的、物种特异性且依赖胚胎阶段的55千道尔顿磷蛋白的定量与特性分析。

Quantitation and characterization of a species-specific and embryo stage-dependent 55-kilodalton phosphoprotein also present in cells transformed by simian virus 40.

作者信息

Chandrasekaran K, McFarland V W, Simmons D T, Dziadek M, Gurney E G, Mora P T

出版信息

Proc Natl Acad Sci U S A. 1981 Nov;78(11):6953-7. doi: 10.1073/pnas.78.11.6953.

Abstract

A 55-kilodalton (kDal) protein was detected recently in primary cultures of day 12 mouse embryos by immunoprecipitation with serum from simian virus 40 (SV40) tumor-bearing hamsters (T serum), Preliminary evidence suggested that this protein was similar to a cellular 55-kDal protein induced after SV40 transformation of mouse cells. We now show that specific approximately 55-kDal [35S]methionine-labeled proteins precipitate from primary cultures of midgestation mouse, rat, and hamster embryos on addition of T serum or monoclonal antiserum prepared against the SV40-induced mouse 55-kDal proteins. The two-dimensional maps of the [35S]methionine-labeled tryptic peptides of the mouse, hamster, and rat embryo proteins are similar to the maps of the corresponding proteins from SV40-transformed cells. Primary cells from midgestation mouse, hamster, or rat embryos contain one-third to one-half as much 55-kDal protein as a SV40-transformed mouse fibroblast cell and nearly the same amount as F9 mouse embryonal carcinoma cells. The amount of 55-kDal protein is greatly reduced on replating the mouse, rat, or hamster embryo primary cells. The amount of this protein in mouse embryos is dependent on the stage of the embryo. The embryo proteins are phosphoproteins.

摘要

最近,通过用来自感染猿猴病毒40(SV40)的仓鼠(T血清)的血清进行免疫沉淀,在第12天小鼠胚胎的原代培养物中检测到一种55千道尔顿(kDal)的蛋白质。初步证据表明,这种蛋白质类似于小鼠细胞经SV40转化后诱导产生的一种细胞55-kDal蛋白质。我们现在表明,在添加T血清或针对SV40诱导的小鼠55-kDal蛋白质制备的单克隆抗血清后,来自妊娠中期小鼠、大鼠和仓鼠胚胎的原代培养物中会沉淀出特定的约55-kDal [35S]甲硫氨酸标记的蛋白质。小鼠、仓鼠和大鼠胚胎蛋白质的[35S]甲硫氨酸标记的胰蛋白酶肽的二维图谱与SV40转化细胞中相应蛋白质的图谱相似。妊娠中期小鼠、仓鼠或大鼠胚胎的原代细胞中55-kDal蛋白质的含量是SV40转化的小鼠成纤维细胞的三分之一到二分之一,与F9小鼠胚胎癌细胞的含量几乎相同。在重新接种小鼠、大鼠或仓鼠胚胎原代细胞后,55-kDal蛋白质的含量大大降低。小鼠胚胎中这种蛋白质的含量取决于胚胎的阶段。胚胎蛋白质是磷蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cda1/349171/cb3c1bd46689/pnas00662-0416-a.jpg

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