MOLECULAR AND FUNCTIONAL STUDIES OF MAXIK CHANNELS

描述(改编自申请者的描述)：这项竞争 续期申请建议进一步研究结构、蜂窝和 Maxik通道功能的遗传机制。主要假设集中在 认为Maxik功能受β亚基联合调节的观点。这个 需要解决的问题包括以下几个方面。1)阿尔法亚单位是如何 结构域在激活过程中移动，Beta亚基对此有何影响 运动？2)酪氨酸激酶如何影响α/β相互作用？3)什么 阿尔法亚单位的结构是C-末端吗？4)这些信号是为了什么 阿尔法亚单位的顶端排序？5)删除 转基因小鼠的α和/或β亚基？拟议的具体目标 是：1)定义阿尔法亚基的不同区域的移动 荧光探针及其由β1亚基修饰。2) 研究了c-Src对α/β复合体的调控机制。3) 对α亚基羧基末端进行大规模纯化 目标是将其具体化。4)研究了α/β亚基的极化分类 在MDCK细胞中。5)获得α和β基因敲除小鼠，并研究 对神经元和血管组织的影响。

DESCRIPTION (adapted from the applicant's description): This competitive renewal application proposes to further examine the structural, cellular and genetic mechanisms of MaxiK channel function. The main hypothesis centers on the idea that maxiK function is regulated by beta subunit association. The questions to be addressed include the following. 1) How do the alpha subunit domains move during activation and how do the beta subunits influence this movement? 2) How do tyrosine kinases influence alpha/beta interaction? 3) What is the structure of the alpha subunit C-terminus? 4) What are the signals for apical sorting of the alpha subunit? 5) What is the impact of deleting the alpha and/or beta subunits from a transgenic mouse? The proposed specific aims are: 1) Define the movement of different regions of the alpha subunit using fluorescent probes, and their modification by the beta 1 subunit. 2) Investigate the mechanism of c-Src modulation of alpha/beta complexes. 3) Perform large-scale purification of the alpha subunit carboxyl end with the goal to crystallize it. 4) Investigate the alpha/beta subunit polarized sorting in MDCK cells. 5) Obtain alpha and beta knockout mice and investigate the impact on neuronal and vascular tissues.