STUDIES OF A NOVEL OXYTOCIN-LIKE PEPTIDE

新型催产素样肽的研究

基本信息

批准号：
3317338
负责人：
JANET A AMICO
金额：
$ 5.55万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-08-01 至 1989-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3317338
关键词：
affinity chromatography blood chemistry estrogens high performance liquid chromatography hormone regulation /control mechanism human tissue neurophysins oxytocin radioimmunoassay radiotracer reversed phase chromatography scintillation spectrometry

项目摘要

In this laboratory, a new protein which is antigenically similar to oxytocin (OT) and arginine vasotocin (AVT) has been found in human plasma and is termed "the novel OT-like peptide." This novel peptide rises in human plasma in response to administration of estrogen (acute and chronic); rises when endogenous levels of estrogen increase (mid-cycle and pregnancy); and is also found in high concentrations in the plasma of patients with chronic renal failure. This novel peptide has heretofore not been described in humans. Reverse phase HPLC separates this material from OT. Two peaks of immunoreactivity, one eluting before and one after OT, are found when plasma from individuals given estrogen, plasma from pregnant women, and plasma from patients with renal failure are chromatographed. All conditions in which the OT-like peptide have been increased have also been characterized by high levels of the estrogen-stimulated neurophysin (ESN), suggesting that ESN and the OT-like peptide are released together and perhaps synthesized and/or are stored together. The quantity of this material in human plasma is less than 10 pg/ml and the identity of this substance is dependent on isolating large amounts of the material. The quantity of plasma needed to recover 100 ng of a salt-free protein - the minimum required for amino acid analysis using the most sensitive of methods - would be in excess of 10 liters. Therefore, a body fluid (as urine from an individual given estrogen or pregnant women or the used dialysate from chronic renal failure patients) and/or a tissue source of the material are being sought and methods of extraction of the peptide, concentration of body fluid and tissue extracts, HPLC isolation and amino acid analysis form one focus of this grant. Because the peptide bears structural similarities to OT, concentrated extracts of the peptide will be tested for oxytocic activity by performing bio- and radioreceptor assays and the release of the OT-like peptide in response to stimuli known to promote release of oxytocin will be tested. By so doing we will learn whether the OT-like peptide and OT are regulated differentially or in parallel and in the process also learn more about the physiology of oxytocin, a hormone about which very little is known.

在这个实验室中，一种与抗原相似的新蛋白质在人血浆中发现了催产素（OT）和精氨酸血管素（AVT）并被称为“新颖的OT型肽”。这个小说的肽在人血浆响应雌激素的给药（急性和慢性）；当雌激素的内源水平增加时（中期和怀孕）;并且在等离子体的高浓度也很高慢性肾衰竭的患者。这个小说肽迄今没有在人类中描述。反向相HPLC将此材料与 OT。两个免疫反应性的峰当来自给予雌激素的个体的血浆，孕妇血浆女性和肾衰竭患者的血浆均具有色谱仪。 OT样肽已增加的所有条件也已以高水平的雌激素刺激的神经素的特征（ESN），表明ESN和OT样肽一起释放也许合成和/或一起存储在一起。这个数量人血浆中的材料小于10 pg/ml，这一点的身份物质取决于隔离大量材料。这恢复100 ng无盐蛋白所需的血浆数量 - 使用最敏感的氨基酸分析所需的最低方法 - 将超过10升。因此，体液（如给予雌激素或孕妇或二手的人的尿液来自慢性肾衰竭患者的透析液）和/或组织源正在寻找材料和提取肽的方法，体液和组织提取物的浓度，HPLC分离和氨基酸分析构成了这笔赠款的一个重点。因为肽熊与OT的结构相似性，肽的浓缩提取物将是通过进行生物和辐射受体测定测试了催产活性并响应已知的刺激而释放OT样肽将测试促进催产素的释放。通过这样做，我们将学习是否对OT样肽和OT进行差异调节，还是在并行和在此过程中还了解有关生理学的更多信息催产素，一种鲜为人知的激素。