High mobility group chromosomal proteins isolated from muclei and cytosol of cultured hepatoma cells are similar.
High mobility group chromosomal proteins isolated from muclei and cytosol of cultured hepatoma cells are similar.
复制标题
从培养的肝癌细胞的细胞核和细胞质中分离出的高迁移率组染色体蛋白是相似的。
DOI：
10.1021/bi00560a013
复制
发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
Bustin,M
中科院分区：
文献类型：
--
作者：
Isackson,PJ;Bidney,DL;Reeck,GR;Neihart,NK;Bustin,M
放大图片作者：Paul J.杰拉尔德·比德尼Reeck，** Natasha K. Neihart和Michael Bustin摘要：使用连续色谱法，在含有固定化双链DNA和单链DNA的柱上，我们从培养的大鼠肝癌细胞的已建立细胞系的胞质溶胶中纯化了一种蛋白质，根据几个标准，该蛋白质是高迁移率族（HMG）蛋白。DNA结合特性、电泳迁移率、氨基酸组成和免疫化学反应性的分析表明，胞质蛋白与从同一细胞系的纯化染色质中分离的HMG-1是相同的蛋白。因此，当哺乳动物细胞用荧光标记的、亲和纯化的抗HMG-1的抗体染色时，真实的HMG-1似乎至少部分负责观察到的细胞质荧光[Bustin，M.，& Neihart，N. K.（1979）Cell 16，181-189]，我们认为HMG-1可以在细胞核和细胞质之间穿梭，可能是响应细胞核对螺旋不稳定蛋白的需要。高迁移率族（HMG）1蛋白是一类非组蛋白染色质蛋白，其可以用0.35M NaCl从染色质中释放并且可溶于2%三氯乙酸（Goodwin et al.，1973年）。术语HMG首先应用于来自小牛胸腺的蛋白质，是指蛋白质在酸-尿素凝胶电泳系统中表现出的高迁移率（Goodwin et al.，1973年）。小牛来自生物化学系，堪萨斯州立大学，曼哈顿，堪萨斯66506（PJI、DLB和GRR），以及分子致癌实验室，国家癌症研究所，国立卫生研究院，贝塞斯达，马里兰州20205（NKH和MB）。1980年4月8日收到。这项工作得到了堪萨斯农业实验站和美国国立卫生研究院（CA-17782）对GRR的资助。来自堪萨斯农业实验站的第80-337-j号贡献。信件应寄给这位作者。他是美国国立卫生研究院研究职业发展奖CA-00425的获奖者。胸腺HMG蛋白具有非常独特的氨基酸组成，具有高含量的酸性和碱性氨基酸残基（Johns等人，1975年）。在遗传多样的生物体中已经发现了在物理、化学或免疫化学性质上与小牛胸腺HMG蛋白相似的蛋白质（沃森等人，1977; Sterner等人，1978; Spiker等人，1978年; Romani等人，1979年）。HMG蛋白首先从染色质中分离出来，因此它们被认为是核组分。然而，最近Bustin和Neihart（1979）提出的证据表明，HMG-1或与HMG-1免疫交叉反应的蛋白质存在于几种培养的哺乳动物细胞的细胞质中以及相同细胞的细胞核中。这一证据是通过显微镜观察
Paul J. Isackson, Dennis L. Bidney, Gerald R. Reeck,** Natasha K. Neihart, and Michael Bustin abstract: Using sequential chromatography on columns containing immobilized double-stranded DNA and single-stranded DNA, we have purified a protein from the cytosol of an establisheo line of cultured rat hepatomacells that, by several criteria, is a high mobility group (HMG) protein. Analyses of DNA binding properties, electrophoretic mobilities, amino acid compositions, and immunochemical re-activities reveal that the cytosolic protein is the same protein as HMG-1 isolated from the purified chromatin of the same cell line. Thus, authentic HMG-1 appears to be at least partially responsible for the cytoplasmic fluorescence observed when mammalian cells are stained with fluorescent-labeled, affinity-purified antibodies against HMG-1 [Bustin, M., & Neihart, N. K.(1979) Cell 16, 181-189], We suggest that HMG-1 can shuttle between nucleusand cytoplasm, perhaps in response to the nucleus’ need for helix destabilizingproteins. e high mobility group (HMG) 1 proteins are a class of nonhistone chromatin proteins that can be released from chromatin with 0.35 M NaCl and that are soluble in 2% trichloroacetic acid (Goodwin et al., 1973). The term HMG, which was first applied to proteins from calf thymus, refers to the high mobility that the proteins exhibit in an acid-urea gel electrophoresis system (Goodwin et al., 1973). The calf tFrom the Department of Biochemistry, Kansas State University, Manhattan, Kansas 66506 (PJI, DLB, and GRR), and the Laboratory of Molecular Carcinogenesis, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20205 (NKH and MB). Received April 8, 1980. This work was supported in part by the Kansas Agricultural Experiment Station and a grant to GRR from the National Institutes of Health (CA-17782). Contribution No. 80-337-j from the Kansas Agricultural ExperimentStation.* Correspondence should be addressed to this author. He is the re-cipient of Research Career DevelopmentAward CA-00425 from the National Institutes of Health. thymus HMG proteins have very distinctive amino acid com-positions with high contents of both acidic and basic amino acid residues (Johns et al., 1975). Proteinsthat are similar in physical, chemical, or immunochemical properties to the calf thymus HMG proteins have been found in phylogenetically diverse organisms (Watson et al., 1977; Sterner et al., 1978; Spiker et al., 1978; Romani et al., 1979). The HMG proteins were first isolated from chromatin, and they have therefore been thought of as nuclear components. Recently, however, Bustin &Neihart (1979) presented evi-dence that HMG-1 or proteins immunologically cross-reactive with HMG-1 occur in the cytoplasm of several types of cul-tured mammalian cells as well as in the nuclei of the same cells. That evidence was obtained by microscopic observations of