Monoclonal antibodies detect conformational abnormality of uracil DNA glycosylase in Bloom's syndrome cells.
Monoclonal antibodies detect conformational abnormality of uracil DNA glycosylase in Bloom's syndrome cells.
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DOI：
10.1093/carcin/8.11.1725
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发表时间：
1987-11
期刊：
影响因子：
4.7
通讯作者：
T. Vollberg;G. Seal;M. Sirover
中科院分区：
文献类型：
--
作者：
T. Vollberg;G. Seal;M. Sirover
用三种抗人胎盘尿嘧啶DNA糖基酶的单抗检测了正常人和Bloom综合征患者尿嘧啶DNA糖基酶的免疫反应性。免疫反应性由三个独立的独立标准确定：酶联免疫吸附试验(ELISA)、酶抑制研究和免疫印迹分析。按各标准定义，正常人尿嘧啶DNA糖基酶与各抗体均呈免疫反应(37.04.12、40.10.09和42.08.07)。相反，从两个单独的未转化的Bloom综合征细胞株中纯化的每个糖基酶都不与抗体40.10.09反应。首先，没有观察到每个糖基酶蛋白的酶联免疫反应。第二，抗体40.10.09不抑制各BLOOM综合征糖基酶的催化作用。然而，每种Bloom综合征酶与抗体37.04.12和42.08.07呈免疫反应。在细胞增殖过程中诱导Bloom综合征酶的过程中，没有观察到有免疫反应的糖基酶种类。免疫印迹分析显示变性Bloom‘s综合征酶与40.10.09抗体发生免疫反应。这些结果表明，Bloom综合征尿嘧啶DNA糖基酶的特征是40.10.09抗体识别的主要抗原部位的天然糖基酶蛋白发生结构变化。这种改变的抗原性可能为鉴定这种人类遗传综合征提供一个免疫学标志。
The immunoreactivity of normal human and Bloom's syndrome uracil DNA glycosylase was examined using a series of three anti-human placental uracil DNA glycosylase monoclonal antibodies. Immunoreactivity was determined by three separate and independent criteria: enzyme-linked immunosorbent assay (ELISA), enzyme inhibition studies and immunoblot analysis. As defined by each criteria, normal human uracil DNA glycosylase was immunoreactive with each antibody (37.04.12, 40.10.09 and 42.08.07). In contrast, each glycosylase purified from two separate non-transformed Bloom's syndrome cell strains was not reactive with antibody 40.10.09. First, no ELISA reactivity was observed with each glycosylase protein. Second, catalysis by each Bloom's syndrome glycosylase was not inhibited by antibody 40.10.09. However, each Bloom's syndrome enzyme was immunoreactive with antibodies 37.04.12 and 42.08.07. No immunoreactive glycosylase species was observed during the induction of the Bloom's syndrome enzyme during cell proliferation. However, immunoreactivity of the denatured Bloom's syndrome enzyme with 40.10.09 antibody was observed by immunoblot analysis. These results suggest that Bloom's syndrome uracil DNA glycosylase is characterized by a structural alteration in the native glycosylase protein secondary to the primary antigenic site recognized by the 40.10.09 antibody. This altered antigenicity may provide an immunological marker for the identification of this human genetic syndrome.