The CRES Gene in Reproduction

生殖中的 CRES 基因

• 批准号: 6776977
• 负责人: Gail A Cornwall
• 金额: $ 9.17万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6776977
• 关键词: SDS polyacrylamide gel electrophoresis; acrosome; clinical research; cysteine; epididymis; gene expression; gene targeting; genetically modified animals; histology; human tissue; immunoprecipitation; in vitro fertilization; laboratory mouse; male reproductive system; polymerase chain reaction; prohormone convertase; proopiomelanocortin; protease inhibitor; protein structure function; serine proteinases; sperm; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): The long-term objective of our research is to determine the role of the CRES (Cystatin-Related Epididymal Spermatogenic) protein in epididymal sperm maturation and sperm function. We have established that CRES defines a new subgroup in the family two cystatins of the cystatin superfamily of cysteine protease inhibitors. Although predicted to structurally resemble the archetype family two cystatins, we have recently demonstrated that CRES does not function as a typical cystatin. We determined that CRES does not inhibit cysteine proteases but rather is a competitive inhibitor (Ki = 25 nM) of the serine protease prohormone convertase 2 (PC2). Because PC2 is a member of a family of proteases with critical roles in the proteolytic maturation of prohormones and proproteins, we propose that CRES may be a new convertase inhibitor that mediates proteolytic processing events important for reproductive function. My short-term goals are to establish whether CRES interacts with PC2 or other convertases in vivo as well as to examine CRES as a convertase inhibitor in greater detail and specifically examine its effects on proprotein processing in cells. Other short-term goals are to examine the biological roles of CRES in vivo including its function in the sperm acrosome and fertilization. My long-term goals are to contribute to our understanding of how proproteins are activated to their mature and functional forms, specifically within spermatozoa and the epididymis, and the mechanisms by which these processes are regulated. Ultimately this may provide mechanisms to target for male contraceptive development. The RCDA, if awarded, will assist me in accomplishing my goals. Specifically, it will release me from Departmental obligations including substantial teaching in medical school gross anatomy, thereby allowing me to devote my full attention to research. In addition, with this award I will not be required to serve on additional committees both at the institutional and departmental levels. The Department of Cell Biology and Biochemistry provides a supportive environment for the development of my research career. Within the department is a large group of NIH-funded reproductive biologists who provide expertise and a collegial, interactive working environment. Also, the newly purchased mass spec will enable on site protein analysis for the studies described above. In short, the environment at TTUHSC is exceptional and will fully support my proposed studies.

描述（由申请人提供）：我们研究的长期目标是确定CRE（胱抑素相关的附子精子生成）蛋白在附睾精子成熟和精子功能中的作用。我们已经确定CRE定义了伴半胱​​氨酸蛋白酶抑制剂的半胱氨酸蛋白酶超家族的两个胱抑素中的一个新亚组。尽管预测在结构上类似于原型家族两个半胱氨酸蛋白酶，但我们最近证明，CRE不起典型的胱抑素的作用。我们确定CRE不会抑制半胱氨酸蛋白酶，而是丝氨酸蛋白酶蛋白酶蛋白酶蛋白酶蛋白酶旋转酶转化酶2（PC2）的竞争抑制剂（Ki = 25 nm）。由于PC2是蛋白酶家族的成员，在蛋白水解成熟中具有关键作用，因此我们建议CRE可能是一种新的转化酶抑制剂，介导对生殖功能重要的蛋白水解处理事件。我的短期目标是确定CRE是否与PC2相互作用或体内其他转化酶相互作用，并更详细地检查CRE作为转化酶抑制剂，并专门检查其对细胞中近亲蛋白加工的影响。其他短期目标是检查CRE在体内的生物学作用，包括其在精子属性和施肥中的功能。我的长期目标是为我们理解如何激活其成熟和功能形式，特别是在精子和附加膜中，以及调节这些过程的机制。最终，这可能为男性避孕发育的目标提供了机制。 RCDA如果被授予，将帮助我实现自己的目标。具体来说，它将使我摆脱部门的义务，包括医学院总体解剖学的大量教学，从而使我全力以赴。此外，有了该奖项，我将不需要在机构和部门一级的其他委员会任职。细胞生物学和生物化学系为我的研究职业的发展提供了一个支持环境。该部门内是一大批由NIH资助的生殖生物学家，他们提供专业知识和合作，互动的工作环境。此外，新购买的质量规格还可以对上述研究进行现场蛋白质分析。简而言之，TTUHSC的环境非常出色，将充分支持我拟议的研究。