Identification of the rat adrenal zona fasciculata/reticularis specific protein, inner zone antigen (IZAg), as the putative membrane progesterone receptor
Identification of the rat adrenal zona fasciculata/reticularis specific protein, inner zone antigen (IZAg), as the putative membrane progesterone receptor
复制标题
DOI：
10.1046/j.1432-1327.2001.02096.x
复制
发表时间：
2001-04-01
期刊：
影响因子：
--
通讯作者：
Vinson, GP
中科院分区：
文献类型：
--
作者：
Raza, FS;Takemori, H;Vinson, GP
使用免疫学方法，一种对大鼠肾上腺皮质内区特异的蛋白质，称为内区抗原（IZAg），先前显示具有26 kDa（IZAg 1）和55-60 kDa（IZAg 2）的两种相互关联的形式，并且对类固醇羟基化具有作用。经双向凝胶电泳和免疫亲和柱纯化后，N-末端氨基酸分析表明，前12个氨基酸与最近描述的假定膜定位孕酮受体（PPMR）的氨基酸相同。RT-PCR Tvas然后用于使用从大鼠肾上腺提取的RNA产生该蛋白质的cDNA。谷胱甘肽S-转移酶（GST）融合构建体在大肠杆菌中表达，并显示产生与其识别为IZAg 1的分子量一致的免疫反应产物。对这种蛋白质的分布进行更详细的检查，不仅在肾上腺皮质的束状核/网状核中，而且在Leydig细胞、肾脏和肝脏中，表明它可能在类固醇激素合成和/或代谢中起作用。
Using immunological methods, a protein specific to the inner zones of the rat adrenal cortex, and called inner zone antigen (IZAg), was previously shown to have two interrelated forms of 26 kDa (IZAg1) and 55-60 kDa (IZAg2), and to have an action on steroid hydroxylation. After two-dimensional gel electrophoresis, and immunoaffinity column purification, N-terminal amino-acid analysis showed that the first 12 amino acids were identical to those of a recently described putative membrane located progesterone receptor (PPMR). RT-PCR Tvas then used to generate the cDNA of this protein, using RNA extracted from rat adrenals. A glutathione S-transferase (GST)-fusion construct was expressed in Escherichia coli, and shown to generate an immunoreactive product of molecular mass consistent with its identification as IZAg1. More detailed examination of the distribution of this protein, not only in the zona fasciculata/reticularis of the adrenal cortex, but also in the Leydig cell, kidney and liver, suggest it may have a role in steroid hormone synthesis and/or metabolism.