Molecular Interactions of the Carboxy Terminus of Prestin

Prestin 羧基末端的分子相互作用

• 批准号: 7199473
• 负责人: ROBERT M RAPHAEL
• 金额: $ 19.64万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7199473
• 关键词: Anions; Auditory; Automobile Driving; Binding; Biochemical; Biological Assay; Carrier Proteins; Cell Volumes; Cell membrane; Cells; Chloride Ion; Chlorides; Co-Immunoprecipitations; Complex; Data; Depth; Diarrhea; Dysplasia; Eukaryota; Eukaryotic Cell; Event; Family; Family member; Fluorescence Recovery After Photobleaching; Fluorescence Resonance Energy Transfer; Gene Family; Hair Cells; Hearing; Helix (Snails); Homeostasis; In Vitro; Ions; Knowledge; Lead; Life; Lipids; Liquid substance; Measures; Mediating; Mediation; Membrane; Membrane Lipids; Membrane Potentials; Membrane Proteins; Molecular; Motor; Mutation; Optics; Organ; Other Finding; Outer Hair Cells; Physiological Processes; Play; Prokaryotic Cells; Property; Proteins; Regulation; Relative (related person); Role; SLC26A3 gene; Specificity; Surface; Syndrome; Techniques; Tertiary Protein Structure; Testing; Work; adenoma; base; ear helix; human disease; insight; member; protein crosslink; protein protein interaction; rat Pres protein; research study; sensor; solute; voltage

DESCRIPTION (provided by applicant): Five years ago, the protein prestin (SLC26A5) was identified as the motor protein that drives electromechanical transduction in cochlear outer hair cells. Subsequent experiments have confirmed prestin is essential for both outer hair cell (OHC) electromotility and normal auditory function. This unique polytopic membrane protein contributes to the voltage sensor that detects changes in the transmembrane potential and to the motor mechanism of OHC electromotility. Prestin is a member of the SLC26A family of anion transporters which play critical roles in ion and fluid homeostasis as well as pH and cell volume regulation. The molecular basis of prestin motor function is presently unknown. The objective of this proposal is to advance our understanding of prestin function. In particular, we will test the hypothesis that prestin molecules self-associate to form functional complexes. The C-terminus of SLC26 family members contains a domain that is highly conserved throughout prokaryotes and eukaryotes, referred to as the STAS domain. Mutations in the STAS domain of several SLC26A proteins are responsible for a variety of human diseases including Pendred syndrome (SLC26A4), congenital chloride diarrhea (SLC26A3), and diastrophic dysplasia (SLC26A2). STAS domains from lower phyla have demonstrated interactions with the membrane. Based on these and other findings, we hypothesize that the STAS domain of prestin mediates prestin-prestin interactions and interactions between prestin and the membrane. Importantly, our preliminary data demonstrates that alterations in the membrane microenvironment alter prestin function. In this proposal, we will use biochemical, cellular, biophysical and optical (FRET and FRAP) approaches to probe the molecular interactions between prestin, prestin and the membrane, and the STAS domain and the membrane. Additionally, we will determine if SLC26A STAS domains can be functionally exchanged. The results from our studies will lead to a deeper understanding, not only of prestin function and the molecular basis of electromotility, but will also provide insights into the function of STAS domains from the SLC26A family.

描述（由申请人提供）：五年前，蛋白质prestin（SLC26A5）被鉴定为驱动人工耳蜗外毛细胞机电转导的运动蛋白。 随后的实验已经证实，Prestin对于外部毛细胞（OHC）电动性和正常听觉功能都是必不可少的。 这种独特的多重膜蛋白有助于电压传感器，该电压传感器检测到跨膜电位变化和OHC电动性的运动机理。 普雷斯汀（Prestin）是SLC26A的阴离子转运蛋白家族的成员，该家族在离子和液体稳态以及pH和细胞体积调节中起关键作用。 Prestin运动功能的分子基础目前尚不清楚。该提案的目的是提高我们对普雷斯汀功能的理解。 特别是，我们将检验以下假设，即普雷斯汀分子会自我关联以形成功能复合物。 SLC26家族成员的C端包含一个在原核生物和真核生物中高度保守的域，称为Stas域。 几种SLC26A蛋白的Stas结构域中的突变负责各种人类疾病，包括Pendred综合征（SLC26A4），先天性氯化物腹泻（SLC26A3）和垂直有性增生不良（SLC26A2）。 来自下属的Stas结构域已证明与膜相互作用。 基于这些发现和其他发现，我们假设普雷斯汀的Stas结构域介导了普雷斯汀 - 普雷斯汀的相互作用以及普雷斯汀和膜之间的相互作用。 重要的是，我们的初步数据表明，膜微环境的改变改变了普雷斯汀功能。在此提案中，我们将使用生化，细胞，生物物理和光学（FRET和FRAP）方法来探测普雷斯汀，普雷斯汀和膜之间的分子相互作用，以及Stas结构域和膜。 此外，我们将确定是否可以在功能上交换SLC26A Stas域。 我们的研究结果将导致更深入地了解Prestin功能和电动性的分子基础，还将提供对SLC26A家族Stas域功能的见解。