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[通过非组蛋白染色质蛋白的³³P磷酸化组分的肝癌相关异源有机肾抗原形成的不可见免疫沉淀物的放射检测]

[Radiometric detection of nonvisualizable immunoprecipitates formed by hepatoma-associated hetero-organic renal antigens of 33P-phosphorylated fractions of non-histone chromatin proteins].

作者信息

Kushner V P, Grandilevskaia A B, Ivanova V A, Fel' V Ia

出版信息

Tsitologiia. 1988 Jan;30(1):53-7.

PMID:3258685
Abstract

A study was made of the antigenic properties of nuclear nonhistone proteins (NHP) from rat tissue eluted with 0.4-0.5 M NaCl gradient on phosphocellulose at very low concentrations. 33P-labeled proteins were obtained by phosphorylation of NHP fractions in the presence of [gamma-33P] ATP. Some non-visual precipitates which appeared during the reaction of specific inhibition of precipitation in agarose gel of 33P-labeled NHP-fractions from rat solid hepatoma 27 cells or rat liver after single hepatocarcinogen diethylnitrosamine injection were detected by means of a calculable radiometric method, proposed by the authors. These precipitates appear only if the rabbit immune sera against NHP-DNA complexes of intact rat kidneys exhausted with liver NHP-antigens was used, never appearing in the case of kidney NHP-antigen exhaustion. Therefore the NHP-fractions prepared from "carcinogenic" liver and rat solid hepatoma cells which possess proper phosphoprotein kinase activity and form specific immune precipitates must be identified as hetero-organic antigens of kidney origin.

摘要

对大鼠组织中核非组蛋白(NHP)的抗原特性进行了研究,这些核非组蛋白是在极低浓度下用0.4 - 0.5M NaCl梯度在磷酸纤维素上洗脱得到的。通过在[γ-33P]ATP存在下对NHP组分进行磷酸化获得33P标记的蛋白质。作者提出的一种可计算的放射测量方法检测到,在单次注射致癌物二乙基亚硝胺后,大鼠实体肝癌27细胞或大鼠肝脏的33P标记NHP组分在琼脂糖凝胶中特异性沉淀抑制反应过程中出现了一些不可见沉淀。这些沉淀仅在使用针对用肝脏NHP抗原耗尽的完整大鼠肾脏的NHP - DNA复合物的兔免疫血清时出现,在用肾脏NHP抗原耗尽的情况下则不会出现。因此,从“致癌性”肝脏和大鼠实体肝癌细胞制备的具有适当磷酸蛋白激酶活性并形成特异性免疫沉淀的NHP组分必须被鉴定为肾脏来源的异源有机抗原。

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