Studies of FSH,FSH-analogues and the FSH-receptor

FSH、FSH 类似物和 FSH 受体的研究

• 批准号: 6988496
• 负责人: JOYCE W LUSTBADER
• 金额: $ 37.52万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6988496
• 关键词: X ray crystallography; carbohydrates; follicle stimulating hormone; glycosylation; hormone analog; hormone receptor; hormone regulation /control mechanism; laboratory mouse; laboratory rat; pharmacokinetics; protein structure; three dimensional imaging /topography

DESCRIPTION (provided by applicant): Modification of the carbohydrate content of hFSH has significant effect on the physiologic properties of the hormone in vivo. To date, preliminary characterization of one such modification has been investigated: hFSH tethered to the CTP from the beta-subunit of hCG (either as a single-chain peptide or as a dimer associated with alpha-subunit). This modification increases the half-life and in vivo bioactivity of the hormone. The effect of fewer or greater numbers of O-linked carbohydrates, the impact on spacing of these moieties, and the potential role of N-linked sugars in this capacity have yet to be thoroughly investigated. These studies will generate important data as to general principles for carbohydrate physiology. By defining the role of differential glycosylation, therapeutic derivatives may be developed through carbohydrate manipulations that may be applicable to all the glycoprotein hormones as well as recombinant proteins in general. In addition, we have preliminary data indicating the feasibility of producing bifunctional hFSH analogues consisting of hFSH tethered to a growth factor. Bifunctionality, both in vitro and in vivo at the level of the ovary was confirmed. This allows for continued and expanded studies on the role of various targeted growth factors on ovarian physiology. Synergistic action on follicular development may also have therapeutic implications for women requiring ovarian stimulation for infertility. Structural knowledge of the interface between FSH and its receptor is essential to developing methods to perturb receptor activity. We plan to determine the structure of the extracellular domain (ECD) of the hFSH receptor, a member of the G protein- coupled receptor superfamily. A three-dimensional representation of the binding region of a glycoprotein hormone receptor will be elucidated for the first time. The structural dynamics and conformational changes in the receptor following ligand binding will provide data crucial to our understanding of the conditions requisite for receptor activation. Together, the studies described in this application will greatly enhance our understanding of gonadotropin biology, the role of various cofactors in ovarian physiology and elicit new methods for altering pharmacodynamics and pharmacokinetics of recombinant proteins.

性状（由申请方提供）：hFSH碳水化合物含量的改变对激素的体内生理特性具有显著影响。迄今为止，已经研究了一种这样的修饰的初步表征：hFSH从hCG的β亚基（作为单链肽或作为与α亚基相关的二聚体）连接到CTP。这种修饰增加了激素的半衰期和体内生物活性。更少或更多数量的O-连接的碳水化合物的影响，对这些部分的间距的影响，以及N-连接的糖在这种能力中的潜在作用还有待深入研究。这些研究将为碳水化合物生理学的一般原理提供重要的数据。通过定义差异糖基化的作用，可以通过碳水化合物操作开发治疗衍生物，其可以适用于所有糖蛋白激素以及一般的重组蛋白。此外，我们有初步的数据表明，生产双功能的hFSH类似物的可行性hFSH拴到生长因子。证实了卵巢水平的体外和体内双功能性。这使得继续和扩大研究的各种靶向生长因子对卵巢生理的作用。对卵泡发育的协同作用也可能对需要卵巢刺激治疗不孕症的妇女有治疗意义。FSH及其受体之间界面的结构知识对于开发干扰受体活性的方法至关重要。我们计划确定hFSH受体（G蛋白偶联受体超家族的成员）的胞外结构域（ECD）的结构。将首次阐明糖蛋白激素受体结合区的三维表示。配体结合后受体的结构动力学和构象变化将为我们理解受体活化所需的条件提供至关重要的数据。总之，本申请中描述的研究将极大地增强我们对促性腺激素生物学、各种辅助因子在卵巢生理学中的作用的理解，并引出改变重组蛋白的药效学和药代动力学的新方法。