FETAL PROTEIN IN CHROMATIN OF NOVIKOFF HEPATOMA AND WALKER 256 CARCINOSARCOMA TUMORS THAT IS ABSENT FROM NORMAL AND REGENERATING RAT-LIVER
FETAL PROTEIN IN CHROMATIN OF NOVIKOFF HEPATOMA AND WALKER 256 CARCINOSARCOMA TUMORS THAT IS ABSENT FROM NORMAL AND REGENERATING RAT-LIVER
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DOI：
10.1073/pnas.73.9.3258
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发表时间：
1976-01-01
影响因子：
11.1
通讯作者：
BUSCH, H
中科院分区：
文献类型：
--
作者：
YEOMAN, LC;JORDAN, JJ;BUSCH, H
Novikoff肝癌细胞染色质蛋白的抗体与Novikoff肝癌、步行者256癌肉瘤和18天胎鼠肝的染色质蛋白在双扩散免疫沉淀试验中形成沉淀带。用于制备抗血清的抗原是最初用3 M NaCl-7 M尿素提取的染色质蛋白，并且在透析至0.14M NaCl-0.35M尿素后可溶。用于分析研究的染色质蛋白用含有0.01 M Tris·HCl（pH 8）和100 μ M苯甲基磺酰氟的0.6 M NaCl提取。正常和18 h再生大鼠肝、心、肾相应的染色质蛋白均未形成沉淀带。用SephadexG-150排阻层析和制备性非变性聚丙烯酰胺凝胶电泳从Novikoff肝癌细胞染色质中纯化抗原。其在变性十二烷基硫酸钠聚丙烯酰胺凝胶上的迁移对应于26，000的MW。氨基酸分析表明，酸性和碱性氨基酸的比例为1.4：1.0。其同质性的证据包括其迁移作为一个单一的蛋白质点的2维聚丙烯酰胺凝胶电泳和其单一的赖氨酸氨基端氨基酸。该蛋白质是一种糖蛋白，每摩尔抗原存在15摩尔半乳糖胺即可证明这一点。这些研究表明，在2个肿瘤的染色质中存在一种胎儿糖蛋白，可能在决定其基因产物中起重要作用。
Antibodies to chromatin proteins of Novikoff hepatoma cells formed precipitin bands in the double diffusion immunoprecipitation assay with chromatin proteins of Novikoff hepatoma, Walker 256 carcinosarcoma and 18 day fetal rat liver. The antigen used for preparation of antiserum was the chromatin proteins initially extracted with 3 M NaCl-7 M urea and soluble after dialysis to 0.14 M NaCl-0.35 M urea. The chromatin proteins used for analytical studies were extracted with 0.6 M NaCl containing 0.01 M Tris.cntdot.HCl (pH 8) and 100 .mu.M phenylmethylsulfonyl fluoride. Corresponding chromatin proteins of normal and 18 h regenerating rat liver, heart and kidney did not form precipitin bands. The antigen was purified from the chromatin of Novikoff hepatoma cells by exclusion chromatography on Sephadex G-150 and preparative nondenaturing polyacrylamide gel electrophoresis. Its migration on denaturing sodium dodecyl sulfate polyacrylamide gels corresponded to a MW of 26,000. Amino acid analysis showed that the ratio of acidic to basic amino acids was 1.4:1.0. Evidence for its homogeneity included its migration as a single protein spot on 2 dimensional polyacrylamide gel electrophoresis and its single lysine amino-terminal amino acid. This protein is a glycoprotein, as shown by the presence of 15 mol of galactosamine per mole of antigen. These studies demonstrate the presence of a fetal glycoprotein in the chromatin of 2 tumors that may have an important role in determining their gene products.