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，一种新的蛋白质，其抗原性与在人血浆中发现了催产素 (OT) 和精氨酸加压素 (AVT) 被称为“新型 OT 样肽”。这种新型肽出现在人血浆对雌激素给药的反应（急性和慢性）；当内源性雌激素水平增加时（周期中期和怀孕）;并且在血浆中也发现高浓度慢性肾功能衰竭患者。这种新颖的肽迄今为止还没有已在人类中被描述。反相 HPLC 将该物质与加时。两个免疫反应峰，一个在 OT 之前洗脱，一个在 OT 之后洗脱，当给予雌激素的个体的血浆、怀孕的血浆时发现女性，并对肾衰竭患者的血浆进行色谱分析。所有 OT 样肽增加的条件也已增加其特点是高水平的雌激素刺激的神经素 (ESN)，表明ESN和OT样肽一起释放并且可能合成和/或存储在一起。这个数量人血浆中该物质的含量低于 10 pg/ml，并且该物质的身份物质依赖于分离大量的材料。这回收 100 ng 无盐蛋白质所需的血浆量 - 使用最灵敏的氨基酸分析所需的最低限度方法 - 将超过 10 升。因此，体液（如服用雌激素的人或孕妇或使用过的人的尿液来自慢性肾功能衰竭患者的透析液）和/或组织来源正在寻找材料和提取肽的方法，体液和组织提取物的浓缩、HPLC 分离和氨基酸分析是本次资助的重点之一。因为肽具有与 OT 结构相似，肽的浓缩提取物将是通过进行生物和放射受体测定来测试催产活性以及响应已知的刺激而释放 OT 样肽将测试促进催产素释放。通过这样做我们将学习 OT样肽和OT是否受到差异调节或在同时，在这个过程中也了解更多关于生理学的知识催产素，一种人们知之甚少的激素。