MOLECULAR AND FUNCTIONAL STUDIES OF MAXIK CHANNELS

MAXIK 通道的分子和功能研究

• 批准号: 6194800
• 负责人: LIGIA G. TORO DE STEFANI
• 金额: $ 34.43万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-17 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6194800
• 关键词: G protein; Xenopus; calcium flux; cellular polarity; chimeric proteins; electrophysiology; fluorescent dye /probe; genetically modified animals; hormone regulation /control mechanism; laboratory mouse; phosphorylation; pore forming protein; potassium channel; protein isoforms; protein kinase; protein protein interaction; protein purification; protein structure function; receptor coupling; site directed mutagenesis; voltage gated channel

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.

描述（根据申请人的描述改编）：此竞争性 更新申请建议进一步检查结构，细胞和 MaxiK通道功能的遗传机制。主要假设集中在 maxiK功能受β亚基联合调节的观点。的 需要解决的问题包括以下方面。1)α亚基是如何 结构域在激活过程中移动，β亚基如何影响它 运动？2)酪氨酸激酶如何影响α/β相互作用？3)什么 是α亚基C末端的结构吗4)这些信号是干什么的 α亚基的顶端排序5)删除 α和/或β亚基的基因吗拟议的具体目标 1）定义α亚基的不同区域的运动，使用 荧光探针以及β 1亚基对其的修饰。（二） 研究α/β复合物的c-Src调节机制。第三章 用以下方法对α亚基羧基末端进行大规模纯化： 4）研究α/β亚基极化分选 在MDCK细胞中。5)获得α和β基因敲除小鼠，并研究 影响神经元和血管组织。