Characterization and isolation of thyroid microsomal antigen.

Characterization and isolation of thyroid microsomal antigen.
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甲状腺微粒体抗原的表征和分离。

DOI:
10.1172/jci112889
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发表时间:
1987
期刊:
The Journal of clinical investigation
影响因子:
--
通讯作者:
DeGroot,LJ
DeGroot,LJ
中科院分区:
--
文献类型:
--
作者:
Hamada,N;Portmann,L;DeGroot,LJ

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我们研究了作为微粒体抗原的107-kD甲状腺蛋白的结构。溶解的微粒体与由IgG组成的亲和凝胶孵育,来自甲状腺炎患者或对照组,与反应凝胶相连。洗脱液采用SDS聚丙烯酰胺电泳和Western blot分析。107- kd和101-kD蛋白在含有患者IgG的凝胶洗脱液中增加,并具有微粒体抗原性。在未还原条件下运行的微粒体的Western blot中,通过抗体鉴定了定义不清的大蛋白。在电泳洗脱和还原条件下重新分析时,它们显示出107-kD抗原。提取还原条件下鉴定的107-kD蛋白,在非还原条件下重新分析。然后观察大分子质量蛋白。双向电泳分离得到一株107-kD抗原,等电点为7.0。微粒体抗原可能是107-kD肽的复合物或多聚体,等电点为7.0
We investigated the structure of the 107-kD thyroid protein recognized as microsomal antigen. Solubilized microsomes were incubated with affinity gels consisting of IgG, from thyroiditis patients or controls, linked to Reacti-gel. Eluates were analyzed by SDS polyacrylamide electrophoresis and Western blot. 107- and 101-kD proteins were augmented in eluates from gels containing patient IgG and had microsomal antigenicity. In a Western blot of microsomes run under unreduced conditions, poorly defined large proteins were identified by antibody. When eluted electrophoretically and reanalyzed in reducing conditions, they demonstrated the 107-kD antigen. The 107-kD protein identified in reducing conditions was extracted and reanalyzed under nonreducing conditions. Large molecular mass proteins were then observed. On two-dimensional electrophoresis, a 107-kD antigen was isolated with isoelectric point of 7.0. The microsomal antigen may be complexes or multimers of a 107-kD peptide with isoelectric point of 7.0.Images
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