K+ channel protein complexes in auditory biology

听觉生物学中的 K 通道蛋白复合物

基本信息

批准号：
7640526
负责人：
WILLIAM R KOBERTZ
金额：
$ 36.56万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7640526
关键词：
Arsenicals Auditory Biochemical Biogenesis Biological Assay Biology Biotinylation Calnexin Cell surface Cells Chemicals Cochlea Cochlear duct Complex Cysteine Data Degradation Pathway Development Disease Ear Endolymph Ensure Extracellular Fluid Face Fractionation Gene Mutation Glycoside Hydrolases Goals Grant Hearing Homeostasis Immunofluorescence Immunologic Intercalated Cell Investigation Jervell-Lange Nielsen Syndrome KCNQ1 protein Knockout Mice Label Laboratories Labyrinth Link Lipids Maps Membrane Metabolic Methods Molecular Mutagenesis Mutation N-terminal Pattern Peptides Physiologic pulse Potassium Potassium Channel Process Property Proteins Reagent Research Research Personnel Retrieval Role Scanning Signal Transduction Site Syncopal Episodes System Testing Time Transmembrane Domain apical membrane cadmium ion congenital deafness crosslink glycosylation hearing impairment inhibitor/antagonist mutant prevent programs protein complex research study trafficking

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this research is to elucidate the molecular and cellular mechanisms that ensure potassium channel protein-protein complexes properly assembly and maintain K+ homeostasis in the cochlear duct. The KCNQ1-KCNE1 K+ channel complex is the exclusive mechanism for endolymphatic K+ secretion into the cochlear duct. Genetic mutations in either KCNQ1 or KCNE that disrupt the assembly, trafficking or function of the complex give rise to Jervell and Lange-Nielsen Syndrome, a recessive form of congenital hearing loss accompanied with syncopal episodes. This application is directed at determining the basic mechanisms of KCNQ1-KCNE1 assembly and trafficking in the inner ear. There are three aims to this application: (1) we will identify the residues that line the protein-protein interface of the KCNQ1-KCNE1 complex utilizing a combination of biochemical and electrophysiological experiments; (2) we will determine the cellular mechanisms that ensure KCNE1 assembles with KCNQ1 by examining the cellular trafficking patterns of the proteins using enzymatic deglycosylation, membrane fractionation, cell surface labeling methods and immunofluorescence; (3) we will investigate a Jervell and Lange-Nielsen Syndrome (JLNS) mutation that disrupts assembly and trafficking of the complex via N-linked glycosylation. For this aim, we will examine the role of N-linked glycosylation in KCNE1 biogenesis, complex assembly and cellular trafficking. The results from these aims will provide a molecular and cellular framework, which will aid in the understanding of JLNS and other KCNQ1-KCNE-linked diseases.

描述（由申请人提供）：这项研究的长期目标是阐明分子和细胞机制，以确保钾通道蛋白 - 蛋白质蛋白复合物适当地组装并维持在人工耳蜗中的K+稳态。 KCNQ1-KCNE1 K+通道复合物是将内淋巴K+分泌到耳蜗管中的专有机制。破坏复合物组装，运输或功能的KCNQ1或KCNE的基因突变会引起Jervell和Lange-Nielsen综合征，这是一种先天性听力损失的隐性形式，并带有张力发作。该应用旨在确定内耳中KCNQ1-KCNE1组装和运输的基本机制。该应用程序有三个目的：（1）我们将使用生物化学和电生理实验的组合来确定KCNQ1-KCNE1复合物的蛋白质蛋白质界面的残基；（2）我们将通过使用酶促脱糖基化，膜分馏，细胞表面标记方法和免疫荧光检查，确定KCNE1通过检查蛋白质的细胞运输模式来确定KCNQ1的细胞机制；（3）我们将研究Jervell和Lange-Nielsen综合征（JLNS）突变，该突变通过N-连接的糖基化破坏了复合物的组装和运输。为此，我们将研究N连接的糖基化在KCNE1生物发生，复杂组装和细胞运输中的作用。这些目标的结果将提供分子和细胞框架，这将有助于理解JLN和其他KCNQ1-KCNE连接疾病。