狼疮血清中自身抗体识别的增殖细胞核抗原的特征分析。
Characterization of proliferating cell nuclear antigen recognized by autoantibodies in lupus sera.
作者信息
Takasaki Y, Fishwild D, Tan E M
出版信息
J Exp Med. 1984 Apr 1;159(4):981-92. doi: 10.1084/jem.159.4.981.
Abstract
A nuclear antigen associated with cell proliferation (proliferating cell nuclear antigen [PCNA]) and blast transformation is recognized by autoantibodies in the sera of some patients with systemic lupus erythematosus. Using this autoantibody as a reagent, PCNA was purified 120-fold by ammonium sulfate fractionation, DEAE chromatography, and Sephadex G200 gel filtration. The antigenicity of PCNA was sensitive to trypsin but resistant to ribonuclease and deoxyribonuclease, suggesting that the antigenic determinant resided in protein and not nucleic acids. PCNA was inactivated at 56 degrees C for 30 min. Isoelectrophoretic focusing showed that the pI was 4.8. Analysis of immunoprecipitates on polyacrylamide gels showed the presence of IgG heavy and light chains and a single polypeptide band of 33,000 mol wt. This polypeptide band was the reactive antigen in immunoblotting (Western transfer) assays.
摘要
一种与细胞增殖相关的核抗原（增殖细胞核抗原[PCNA]）和母细胞转化可被一些系统性红斑狼疮患者血清中的自身抗体识别。以这种自身抗体作为试剂，通过硫酸铵分级分离、DEAE柱层析和Sephadex G200凝胶过滤，PCNA被纯化了120倍。PCNA的抗原性对胰蛋白酶敏感，但对核糖核酸酶和脱氧核糖核酸酶有抗性，这表明抗原决定簇存在于蛋白质而非核酸中。PCNA在56℃下30分钟即失活。等电聚焦显示其pI为4.8。对聚丙烯酰胺凝胶上的免疫沉淀物进行分析，结果显示存在IgG重链和轻链以及一条分子量为33,000的单一多肽带。这条多肽带是免疫印迹（Western印迹）分析中的反应性抗原。
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