STRUCTURE & FUNCTION OF THE EPIDIDYMIS AND VAS DEFERENS

结构

• 批准号: 6520746
• 负责人: DAVID W HAMILTON
• 金额: $ 20.88万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-29 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520746
• 关键词: SDS polyacrylamide gel electrophoresis; acrosome; calcium channel; calcium flux; confocal scanning microscopy; cysteine; epididymis; fertilization; immunoprecipitation; laboratory mouse; laboratory rat; membrane fusion; polymerase chain reaction; protein binding; protein structure; sperm; sperm capacitation; western blottings

Funds are requested to carry out projects on the function of proteins of the Crisp superfamily in the male reproductive tract. Three Crisp genes encode proteins in an organ-specific manner: Crisp-2 is testis- specific, Crisp-3 is submandibular gland-specific and Crisp-1 is expressed primarily in the epididymis. Crisp genes are found in mammals as diverse as rats, guinea pigs, mice and humans and so must have an important function in the organs within which they are expressed. Crisp-1 proteins are synthesized and secreted by the epididymal epithelium and subsequently bind to sperm in a domain-specific manner in rats. Protein D, which binds both loosely and tightly, localizes to the head of the sperm on the plasma membrane overlying the acrosome and has been shown to play a role in fertilization. Protein E binds specifically to plasma membrane of the tail, and also participates in fertilization. Proteins D and E have significant homology to helothermine (a salivary gland toxin that blocks ryanodine receptors) and to a number of insect and snake venom proteins that are capable of regulating ion channels. We hypothesize that proteins D and E function to regulate sperm ion fluxes, particularly calcium. In the first specific aim we explore the roles of Crisp-1 during sperm-egg fusion. In the second aim, we explore their role in capacitation and the acrosome reaction, both processes that require large ion fluxes. Finally, in the last specific aim we address the question of mechanism(s) that mediate binding of protein D and E to sperm in the epididymis. We hypothesize that the signal for domain localization resides in the NH2 terminus of the molecule and the carboxyl terminus, where 14 of the 16 cysteines are found, provides the plasma membrane binding domain. These studies will contribute significant knowledge on the role of a potentially important group of proteins both in the physiology of the epididymis and in fertilization.

要求提供资金以开展关于蛋白质功能的项目 男性生殖道中的脆蛋白超家族三声清脆 基因以器官特异性方式编码蛋白质：Crisp-2是睾丸- 具体地，Crisp-3是下颌下腺特异性的，而Crisp-1是下颌下腺特异性的。 主要在附睾中表达。在哺乳动物中发现了脆的基因 如大鼠、豚鼠、小鼠和人类， 在表达它们的器官中发挥重要作用。 Crisp-1蛋白由附睾合成和分泌 并随后以结构域特异性方式与精子结合 对大鼠蛋白质D，结合松散和紧密，定位于 精子头部位于顶体上的质膜上， 已经被证明在受精过程中起作用。 蛋白E结合 特别是尾部的质膜，并且还参与 受精 蛋白质D和E具有显著的同源性， helothermine（一种阻断ryanodine受体的唾液腺毒素） 以及许多昆虫和蛇毒蛋白， 调节离子通道。 我们假设蛋白质D和E的功能 来调节精子离子流，特别是钙离子。 上 具体目的是探讨Crisp-1在精卵融合过程中的作用。 在第二个目标中，我们探讨了它们在获能中的作用， 顶体反应，这两个过程都需要大的离子通量。 最后，在最后一个具体目标中，我们解决了以下问题： 调节蛋白D和E与精子结合的机制 附睾 我们假设用于域定位的信号 位于分子的NH 2末端和羧基末端， 其中发现16个半胱氨酸中的14个，提供质膜 结合域 这些研究将有助于重要的知识， 一组潜在的重要蛋白质在 附睾生理学和受精。