STRUCTURE/FUNCTION OF A PREGNANCY ASSOCIATED PROTEIN

怀孕相关蛋白质的结构/功能

• 批准号: 6042742
• 负责人: THOMAS ROSS HANSEN
• 金额: $ 15.87万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6042742
• 关键词: complementary DNA; confocal scanning microscopy; cow; endometrium; female; fluorescence microscopy; gene expression; genetic transcription; immunoaffinity chromatography; immunoconjugates; interferon gamma; laboratory mouse; laboratory rabbit; pregnancy; pregnancy immunology; protein sequence; protein structure function; sex cycle; sheep; species difference; synthetic peptide; tissue /cell culture; ubiquitin

A subst of interferon (IFN) stimulated genes (ISG) encodes human 15- kDa (ISG15) and bovine 17-kDa (ISG17) ubiquitin homologs that share similar structure and function. ISG15 is induced in several cell lines in response to IFN and in decidualized human stromal cells in response to pregnancy. ISG17, however, appears to be primarily induced in the bovine endometrium in response to conceptus-derived IFN-tau. These ISGs conjugate to intracellular proteins in response to IFN. The fate and identify of proteins conjugated to these ISGs have not been determined. ISG17 is realized by the endometrium from day 15 to 26 of pregnancy in cows and in endometrial cells treated with IFN-tau. Also, ISG15 has immunomodulatory roles, an example of which is induction of release of IFN-gamma. It is propose to use a bovine model to study the intra- and extra-cellular function of these ISGs. Recombinant (r) ISG17 will be generated using a yeast expression system, purified, and evaluated for bioactivity/function using in vitro and in vivo approaches. Proteins that become conjugated to ISG17 in response to IFN-tau and pregnancy will be identified using antibody-based affinity, molecular biology, and hypothesis-based approaches. Finally, it is proposed to determine tissue distribution and cytosolic trafficking of ISG17 and its conjugates in endometrium during pregnancy and in endometrial cells in response to IFN-tau using epifluorescent and confocal microscopy.

干扰素（IFN）刺激基因（ISG）的替代物编码人类15-KDA（ISG15）和牛17-KDA（ISG17）泛素同源物，它们具有相似的结构和功能。 ISG15在响应于妊娠的IFN和de虫的人类基质细胞中响应于几个细胞系中诱导。然而，ISG17似乎主要是在牛子宫内膜中诱导的，这是响应于概念的IFN-TAU而引起的。这些ISG响应于IFN响应于细胞内蛋白。尚未确定与这些ISG结合的蛋白质的命运和识别。 ISG17是通过子宫内膜从奶牛和用IFN-TAU处理的子宫内膜细胞中的怀孕和子宫内膜细胞中实现的。同样，ISG15具有免疫调节作用，其中IFN-gamma释放的一个例子。建议使用牛模型研究这些ISG的细胞内和细胞外功能。重组（R）ISG17将使用酵母表达系统生成，使用体外和体内方法进行纯化并评估生物活性/功能。将使用基于抗体的亲和力，分子生物学和基于假设的方法来鉴定与IFN-TAU和妊娠相结合的蛋白质。最后，建议在怀孕期间确定ISG17及其在子宫内膜中的结合物和子宫内膜细胞中的组织分布和胞质运输，并使用落叶和共斑显微镜响应IFN-TAU。