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）的一个亚群编码具有相似结构和功能的人15- kDa（ISG 15）和牛17-kDa（ISG 17）泛素同源物。ISG 15在几种细胞系中响应于IFN而被诱导，并且在蜕膜化的人基质细胞中响应于妊娠而被诱导。然而，ISG 17似乎主要在牛子宫内膜中响应于孕体来源的IFN-tau而被诱导。这些ISG响应于IFN而与细胞内蛋白质缀合。尚未确定与这些ISG缀合的蛋白质的命运和鉴定。ISG 17由母牛妊娠第15 - 26天的子宫内膜和用IFN-tau处理的子宫内膜细胞实现。此外，ISG 15具有免疫调节作用，其实例是诱导IFN-γ的释放。建议使用牛模型来研究这些ISG的细胞内和细胞外功能。重组（r）ISG 17将使用酵母表达系统生成，纯化，并使用体外和体内方法评价生物活性/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响应于IFN-tau和妊娠而与ISG 17缀合的蛋白质。最后，建议使用落射荧光显微镜和共聚焦显微镜来确定ISG 17及其缀合物在妊娠期间子宫内膜中以及在子宫内膜细胞中响应于IFN-tau的组织分布和胞质运输。