Structure-function analysis of Mullerian Inhibiting Substance (MIS)

项目摘要：苗勒管抑制物质（MIS）或抗苗勒管激素（AMH），最初确定 由于其在发育过程中的雄性性别分化中的作用，已成为雌性发育中的重要分子。 生殖例如，MIS在调节卵泡发育和血清水平中起重要作用， 现在被用来衡量卵巢储备。MIS中的突变与男性和女性都相关 生殖系统疾病，包括男性持续性苗勒管综合征（PMDS）和多囊卵巢 女性多囊卵巢综合征（PCOS）。作为TGFβ家族的一员，MIS通过I型和II型受体进行信号传导。 独特的是，MIS通过其自身的II型受体MISRII和I型受体Alk2发出信号， 多配体。虽然以前的研究详细说明了TGFβ家族配体的相互作用，但MIS如何相互作用， 分子水平，MISRII和Alk2是未知的。本提案的目的是了解MIS如何 与其同源受体相互作用以及如何通过这些受体产生特异性MIS信号传导。在这里， 我们将联合收割机结合结构和功能的方法，包括X射线晶体学，基于细胞的信号传导， 测定、基于结构的工程以及生育力和卵巢保护的生物模型。我们将继续 以下三个具体目标：（1）用MISRII和Alk 2确定MIS的晶体结构，（2）表征MIS的晶体结构。 MIS受体，并确定如何产生MIS特异性信号，使用细胞外和 （3）产生MIS类似物，其将在体外结合和基于细胞的结合中进行测试。 沿着体内测定，包括卵泡抑制的AAV 9模型。总的来说，这项建议将 揭示MIS如何与其受体相互作用，为开发修饰MIS活性的试剂提供平台 具有未来在生殖治疗中应用的潜力。

Project Summary: Müllerian Inhibiting Substance (MIS) or Anti- Müllerian Hormone (AMH), originally identified for its role in male sex differentiation during development, has emerged as a significant molecule in female reproduction. For example, MIS plays an important role in regulating follicle development and serum levels and is now used as a measure of ovarian reserve. Mutations in MIS are associated with both male and female reproductive disorders, including Persistent Müllerian Duct Syndrome (PMDS) in males and Polycystic Ovary Syndrome (PCOS) in females. As a member of the TGFβ family, MIS signals through a type I and type II receptor. Uniquely, MIS signals through its own type II receptor MISRII and the type I receptor Alk2, which is utilized by multiple ligands. While previous studies have detailed TGFβ family ligand interactions, how MIS interacts, at the molecular level, with MISRII and Alk2 is unknown. The objective of this proposal is to understand how MIS interacts with its cognate receptors and how specific MIS signaling is generated through these receptors. Here, we will combine both structural and functional approaches, including X-ray crystallography, cell-based signaling assays, structure-based engineering, and biological models of fertility and ovarian protection. We will pursue the following three specific aims (1) determine the crystal structure of MIS with MISRII and Alk2, (2) characterize the MIS receptors and determine how the generate MIS-specific signaling, using both the extracellular and intercellular domains and (3) generate MIS analogs that will be tested in both in vitro binding and cell-based assays, along with in vivo assay including an AAV9 model of follicle suppression. Collectively, this proposal will uncover how MIS interacts with its receptors providing a platform for developing reagents that modify MIS activity with the potential for future application in reproductive therapies.