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首页> 外文期刊>Molecular and Cellular Biology >Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes.
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Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes.

机译：失活的孕激素受体复合物的新型23-千达尔顿蛋白的表征。

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Immunoprecipitation of unactivated avian progesterone receptor results in the copurification of hsp90, hsp70, and three additional proteins, p54, p50, and p23. p23 is also present in immunoaffinity-purified hsp90 complexes along with hsp70 and another protein, p60. Antibody and cDNA probes for p23 were prepared in an effort to elucidate the significance and function of this protein. Antibodies to p23 detect similar levels of p23 in all tissues tested and cross-react with a protein of the same size in mice, rabbits, guinea pigs, humans, and Saccharomyces cerevisiae, indicating that p23 is a conserved protein of broad tissue distribution. These antibodies were used to screen a chicken brain cDNA library, resulting in the isolation of a 468-bp partial cDNA clone encoding a sequence containing four sequences corresponding to peptide fragments isolated from chicken p23. This partial clone was subsequently used to isolate a full-length human cDNA clone. The human cDNA encodes a protein of 160 amino acids that does not show homology to previously identified proteins. The chicken and human cDNAs are 88% identical at the DNA level and 96.3% identical at the protein level. p23 is a highly acidic phosphoprotein with an aspartic acid-rich carboxy-terminal domain. Bacterially overexpressed human p23 was used to raise several monoclonal antibodies to p23. These antibodies specifically immunoprecipitate p23 in complex with hsp90 in all tissues tested and can be used to immunoaffinity isolate progesterone receptor complexes from chicken oviduct cytosol.
机译：未激活的禽孕激素受体的免疫沉淀作用导致hsp90，hsp70和其他三种蛋白质p54，p50和p23的共纯化。 p23与hsp70和另一种蛋白质p60一起也存在于免疫亲和纯化的hsp90复合物中。制备p23的抗体和cDNA探针是为了阐明该蛋白的重要性和功能。 p23抗体在所有测试的组织中检测到相似水平的p23，并与小鼠，兔子，豚鼠，人类和酿酒酵母中相同大小的蛋白质发生交叉反应，表明p23是广泛组织分布的保守蛋白质。这些抗体用于筛选鸡脑cDNA文库，从而分离出468 bp的部分cDNA克隆，该克隆编码一个序列，该序列包含四个与从鸡p23分离的肽片段相对应的序列。该部分克隆随后用于分离全长人cDNA克隆。人cDNA编码一个160个氨基酸的蛋白质，该蛋白质与以前鉴定的蛋白质没有同源性。鸡和人的cDNA在DNA水平上相同，分别为88％和96.3％。 p23是一种高酸性磷酸蛋白，具有富含天冬氨酸的羧基末端结构域。细菌过度表达的人p23被用来产生几种针对p23的单克隆抗体。这些抗体可在所有测试的组织中与hsp90特异性免疫沉淀p23，可用于从鸡输卵管细胞溶胶中免疫亲和分离孕激素受体复合物。

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