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) 被鉴定为驱动耳蜗外毛细胞机电转导的运动蛋白。 随后的实验证实 prestin 对于外毛细胞 (OHC) 电动性和正常听觉功能至关重要。 这种独特的多面体膜蛋白有助于检测跨膜电位变化的电压传感器和 OHC 电动性的运动机制。 Prestin 是阴离子转运蛋白 SLC26A 家族的成员，在离子和液体稳态以及 pH 和细胞体积调节中发挥着关键作用。 prestin 运动功能的分子基础目前尚不清楚。该提案的目的是增进我们对 prestin 功能的理解。 特别是，我们将测试 prestin 分子自缔合形成功能复合物的假设。 SLC26家族成员的C末端含有一个在原核生物和真核生物中高度保守的结构域，称为STAS结构域。 几种 SLC26A 蛋白 STAS 结构域的突变导致多种人类疾病，包括 Pendred 综合征 (SLC26A4)、先天性氯化物腹泻 (SLC26A3) 和萎缩性发育不良 (SLC26A2)。 来自较低门的 STAS 结构域已被证明与膜存在相互作用。 基于这些和其他发现，我们假设 prestin 的 STAS 结构域介导 prestin-prestin 相互作用以及 prestin 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会改变 prestin 功能。在本提案中，我们将使用生物化学、细胞、生物物理和光学（FRET 和 FRAP）方法来探测 prestin、prestin 和膜以及 STAS 结构域和膜之间的分子相互作用。 此外，我们将确定 SLC26A STAS 域是否可以进行功能交换。 我们的研究结果不仅将带来对 prestin 功能和电动性分子基础的更深入理解，还将提供对 SLC26A 家族 STAS 结构域功能的深入了解。