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）：本研究的长期目标是阐明确保钾通道蛋白-蛋白复合物正确组装并维持耳蜗管中 K+ 稳态的分子和细胞机制。 KCNQ1-KCNE1 K+ 通道复合体是内淋巴 K+ 分泌到耳蜗管的唯一机制。 KCNQ1 或 KCNE 中的基因突变会破坏复合体的组装、运输或功能，从而导致杰维尔和兰格-尼尔森综合征，这是一种伴有晕厥发作的隐性先天性听力损失。该应用旨在确定 KCNQ1-KCNE1 在内耳中组装和运输的基本机制。该应用有三个目标：（1）我们将利用生化和电生理学实验的结合来识别 KCNQ1-KCNE1 复合物的蛋白质-蛋白质界面上的残基； (2) 我们将通过酶促去糖基化、膜分级分离、细胞表面标记方法和免疫荧光检查蛋白质的细胞运输模式，确定确保 KCNE1 与 KCNQ1 组装的细胞机制； (3) 我们将研究 Jevell 和 Lange-Nielsen 综合征 (JLNS) 突变，该突变通过 N 连接糖基化破坏复合物的组装和运输。为此，我们将研究 N 连接糖基化在 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一个分子和细胞框架，这将有助于理解 JLNS 和其他 KCNQ1-KCNE 相关疾病。