'REGULATORY MECHANISMS OF KALIRIN'S GEF1 DOMAIN'

描述(由申请人提供)： 了解神经元如何启动轴突生长非常重要，而不仅仅是对 我们对神经系统和神经元发育的基本认识 形态学，但也有助于神经学疗法的发展 精神错乱。我的长期目标是了解 鸟嘌呤核苷酸交换因子和GTP酶调节神经元形态， 作为使用卡利林辅助神经元再生的基础 战略。这个RCDA的目标是让我获得更多 专业知识，培养一名学术科学家所需的几项技能， 在一个新的研究领域发展一个历史，并开始建立一个新的 研究计划。在这项提议的过程中，一个多方面的发展 议程将包括参加和运营期刊俱乐部，发表演讲， 上课，授课，接受优质导师相约 这些发展目标。我将在多维核磁共振方面得到Dr。 格伦·金，一位工作效率极高的核磁共振光谱专家 唱片。贝蒂·艾珀博士将帮助我培养管理 新建实验室，授课，培训员工。埃珀博士已经做了很多 对神经肽领域的贡献，并具有悠久的历史 培养未来的科学领袖。几项研究和我们的初步研究 实验表明，Kalirin在轴突的形成和分化中起关键作用。 神经元分化。考虑孤立加里林的稳健效应 GEF1领域对神经元发育的影响，本提案的目的是为了更好地 理解完整的加里林的GEF1结构域是通过什么机制(S) 受监管的。我们将首先确定加里林的结构域(S)，它调节 GEF1结构域通过产生一系列突变型缺失Kalirin蛋白和 在轴突生长和全球环境基金激活试验中对它们进行测试。使用 从这些实验中获得的信息，我们将从几个 已确定的监管结构域可能控制的潜在机制 GEF1域。最后，机械论假说将在 用核磁共振法测定分子水平的三维结构 GEF1及其调控元件的研究和配体的作用 结构上的相互作用。了解全球环境基金 加里林的区域被控制将为未来的研究提供基础 确定Kalirin控制神经元形态的机制。

DESCRIPTION (provided by applicant): Understanding how neurons initiate neurite outgrowth is important, not just for our basic understanding of development of the nervous system and neuronal morphology, but also to aid in the development of therapies for neurological disorders. My long-term goal is to understand the molecular mechanisms by which guanine nucleotide exchange factors and GTPases modulate neuronal morphology, as a basis for using Kalirin to aid in the development of neuronal regeneration strategies. The goal of this RCDA is to allow me to obtain additional expertise, cultivate several of the skills necessary for an academic scientist, develop a history in a new field of study, and begin to establish a new research program. During the course of this proposal a multifaceted development agenda will involve attending and running journal clubs, giving presentations, taking classes, teaching classes and receiving high-quality mentorship to meet these developmental goals. I will be mentored in multidimensional NMR by Dr. Glenn King, an expert NMR spectroscopist with an outstanding productivity record. Dr. Betty Eipper will help me cultivate the skills needed to manage a new laboratory, teach classes, and train staff. Dr. Eipper has made many seminal contributions to the neuropeptide field and has a long-history of training future leaders in science. Several studies and our preliminary experiments indicate that Kalirin plays a key role in neurite formation and neuronal differentiation. Considering the robust effect of the isolated kalirin GEF1 domain on neuronal development, the goal of this proposal is to better understand the mechanism(s) by which the GEF1 domain of intact kalirin is regulated. We will first identify the domain(s) of kalirin that regulate the GEF1 domain by generating a series of mutant deletion kalirin proteins and testing them in neurite outgrowth and GEF activation assays. Using the information gained from these experiments, we will pursue one of several potential mechanisms by which the identified regulatory domain might control the GEF1 domain. Finally, the mechanistic hypotheses will be further refined at the molecular level by using NMR to determine the three-dimensional structure of the GEF1 with its regulatory elements and examining the effects of ligand interactions upon the structure. Understanding the mechanism by which the GEF domain of kalirin is controlled will provide a basis for future studies to determine the mechanisms by which kalirin controls neuronal morphology.