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[3H]苯并(a)芘代谢物与AKR小鼠胚胎细胞系核蛋白的结合。

Binding of [3H]benzo(a)pyrene metabolites to the AKR mouse embryo cell line nuclear proteins.

作者信息

Zytkovicz T H, Moses H L, Spelsberg T C

出版信息

Cancer Res. 1981 May;41(5):1608-14.

PMID:6260346
Abstract

Nuclear proteins from AKR-2B mouse embryo cells incubated 24 hr with [3H]benzo(a)pyrene (1 microgram/ml; 10 microCi/ml) were purified to remove nucleic acids and characterized by isoelectric focusing and sodium dodecyl sulfate gel electrophoresis on polyacrylamide gels. Analysis of the nuclear proteins by isoelectric focusing revealed a distribution of protein in the pH gradient with two prominent bands of proteins, one focusing at pH 5 and the other at pH 11, both of which bound significant amounts of the carcinogen. The acid-soluble proteins from the nuclear preparation were analyzed by isoelectric focusing, and the results also revealed two peaks of protein and radioactivity at pH's of 5 and 11. Further analysis of the acid-soluble fraction on acid urea gels revealed that significant radioactivity was associated with proteins that migrated near but slower than histone H1 which may be the high-mobility group proteins 1 and 2. Radioactivity also comigrated with histones H3 and H2B. Low levels of radioactivity were associated with histone H1 and H4.

摘要

将AKR - 2B小鼠胚胎细胞与[3H]苯并(a)芘(1微克/毫升;10微居里/毫升)一起孵育24小时后,提取细胞核蛋白,对其进行纯化以去除核酸,并通过等电聚焦和聚丙烯酰胺凝胶上的十二烷基硫酸钠凝胶电泳进行表征。通过等电聚焦分析细胞核蛋白,结果显示蛋白质在pH梯度中的分布情况,有两条明显的蛋白带,一条聚焦在pH 5,另一条聚焦在pH 11,这两条带都结合了大量的致癌物。对细胞核制剂中的酸溶性蛋白进行等电聚焦分析,结果也显示在pH 5和11处有两个蛋白和放射性峰。在酸性尿素凝胶上对酸溶性部分进行进一步分析发现,大量放射性与迁移速度比组蛋白H1稍慢且靠近H1的蛋白相关,这些蛋白可能是高迁移率族蛋白1和2。放射性也与组蛋白H3和H2B共迁移。低水平的放射性与组蛋白H1和H4相关。

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