STRUCTURE/FUNCTION OF A PREGNANCY-ASSOCIATED PROTEIN

怀孕相关蛋白质的结构/功能

• 批准号: 2205582
• 负责人: THOMAS ROSS HANSEN
• 金额: $ 8.29万
• 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 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2205582
• 关键词: antiserum; cell type; complementary DNA; computer assisted sequence analysis; cow; endometrium; gene expression; genetic regulation; genetic transcription; high performance liquid chromatography; immunoaffinity chromatography; in situ hybridization; laboratory rabbit; northern blottings; pregnancy; protein purification; protein sequence; protein structure function; proteins; sex cycle; southern blotting; species difference; synthetic peptide; tissue /cell culture; western blottings

The long-term objectives of the research ar to understand the function, regulation, and mechanism of action of a 16 kDa protein that is secreted in high amounts during pregnancy by the bovine uterine endometrium. The function of this protein has implications in several mammalian species, including humans, since it shares epitopes with ubiquitin and a 15-kDa ubiquitin cross-reactive protein (UCRP), and is regulated by conceptus- derived interferon tau (IFN-). Thus, this protein may serve as an index of early pregnancy and potentially, fertility. To meet the long-term objectives of the research, we intend to focus on the elucidation of structure and pregnancy-associated expression of the 16-kDa protein. Specifically, amino-terminal and internal peptide amino acid sequence will be determined following purification by using fast phase liquid chromatography. The cDNA(s) encoding the 16-kDa protein will be sequenced and used in Northern blots and in situ hybridization to confirm pregnancy-associated transcription. The number, complexity and distribution of genes across species will be studied by using Southern blots. A more specific polyclonal antiserum will be prepared and used to detect the 16-kDa protein in uterine secretions and serum. It also will be used to make an immunoaffinity matrix that will aid in purification. Immunosuppressive activity will be determined in rosette inhibition and anti-proliferative tests. Future experiments, by using cDNAs to generate recombinant 16-kDa protein, will be directed towards identifying function and mechanism of action during early pregnancy. Study of this 16-kDa protein has potential implication in human fertility, since communication between the conceptus and uterus is similar across many mammalian species.

研究的长期目标是了解功能， 调节和作用机制的16 kDa的蛋白质，分泌在 在怀孕期间，牛子宫内膜分泌大量的蛋白质。 的 这种蛋白质的功能在几种哺乳动物中有意义， 包括人类，因为它与泛素和15-kDa的 泛素交叉反应蛋白（UCRP），并受孕- 衍生的干扰素tau（IFN-）。 因此，这种蛋白质可以作为一种指标， 早孕以及潜在的生育能力。 为达致研究的长远目标，我们打算集中研究 16-kDa的结构和妊娠相关表达的阐明 蛋白 具体地，氨基末端和内部肽氨基酸 在纯化后通过使用快速相 液相色谱法 编码16-kDa蛋白的cDNA将被纯化。 测序并用于北方印迹和原位杂交以确认 妊娠相关转录。 数量、复杂性和 跨物种的基因分布将通过使用Southern杂交来研究。 污点。 将制备更具特异性的多克隆抗血清，并用于 检测子宫分泌物和血清中的16-kDa蛋白。 还应当 用于制备有助于纯化的免疫亲和基质。 免疫抑制活性将在玫瑰花结抑制中测定， 抗增殖试验 未来的实验，通过使用cDNA产生 重组16-kDa蛋白，将针对鉴定功能 和作用机制。 16-kDa的研究 蛋白质在人类生育中具有潜在的意义， 在许多哺乳动物物种中，孕体和子宫之间的关系是相似的。