Structural correlates of prestin activity.

描述(申请人提供)：随着prestin被鉴定为外毛细胞(OHC)难以捉摸的侧膜马达蛋白，我们面临着了解这种单一分子如何影响哺乳动物精致听力的可能性。为此，我们的兴趣集中在确定什么蛋白质结构可能引起马达的已知生物物理属性，包括它的温度、张力和电压依赖性。我们假设这些分子活性的产生和/或受到Prestin细胞内C和N末端与其他细胞内蛋白质和阴离子的相互作用的影响，也可能受到多聚体相互作用的影响。我们建议针对一组目标，包括1)确定prestin的C和N末端对prestin标志性生物物理属性的贡献，2)确定prestin在我们开发的prestin细胞系中的运输途径，以及3)确定prestin(SLC26A5)与其邻近家族成员slc26a6之间的哪些结构不同，从而解释prestin的非线性电容。为了达到这些目标，我们将采用一系列电生理、分子生物学和生物化学方法。我们相信，通过这些研究获得的信息将有助于理解OHC如何使我们能够如此清晰地听到声音，进而我们如何与OHC折磨数百万人的病理作斗争。

DESCRIPTION (provided by applicant): With the identification of prestin as the elusive lateral membrane motor protein of the outer hair cell (OHC), we are faced with the possibility of understanding how this single molecule can affect the mammal's exquisite sense of hearing. To that end, we have focused our interest on determining what protein structures may give rise to the motor's known biophysical attributes, including its temperature, tension, and voltage dependence. We hypothesize that these molecular activities arise and/or are influenced by interactions of prestin's intracellular C and N termini with other intracellular proteins and anions, and possibly by multimeric interactions, as well. We propose to target a focused set of aims, including 1) determine the contribution of prestin's C and N termini to prestin's signature biophysical attributes, 2) determine prestin's trafficking route in a prestin cell line that we have developed, and 3) determine what structures are different between prestin (slc26a5) and its closet family member slc26a6 that account for prestin's nonlinear capacitance. In order to reach these goals, we will employ a host of electrophysiological, molecular biological and biochemical methods. We believe that the information that we obtain through these studies will aid in understanding how the OHC enables us to hear so well and in turn how we might combat pathologies of the OHC that afflict millions.