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IR Laser-Induced Perturbations of the Voltage-Dependent Solute Carrier Protein SLC26a5

机译:红外激光诱导的电压依赖性溶质载体蛋白SLC26a5的扰动

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摘要

Alterations in membrane capacitance can arise from linear and nonlinear sources. For example, changes in membrane surface area or dielectric properties can modify capacitance linearly, whereas sensor residues of voltage-dependent proteins can modify capacitance nonlinearly. Here, we examined the effects of fast temperature jumps induced by an infrared (IR) laser in control and prestin (SLC26a5)-transfected human embryonic kidney (HEK) cells under whole-cell voltage clamp. Prestin’s voltage sensor imparts a characteristic bell-shaped, voltage-dependent nonlinear capacitance (NLC). Temperature jumps in control HEK cells cause a monophasic increase in membrane capacitance (Cm) regardless of holding voltage due to double-layer effects. Prestin-transfected HEK cells, however, additionally show a biphasic increase/decrease in Cm with a reversal potential corresponding to the voltage at peak NLC of prestin (Vh), attributable to a rapid temperature-following shift in Vh, with shift rates up to 14 V/s over the course of a 5 ms IR pulse. Treatment with salicylate, a known inhibitor of NLC, reestablishes control cell behavior. A simple kinetic model recapitulates our biophysical observations. These results verify a voltage-dependent protein’s ability to respond to fast temperature perturbations on a par with double-layer susceptibility. This likely arises from prestin’s unique ability to move sensor charge at kilohertz rates, which is required for the outer hair cells’ role as a cochlear amplifier.
机译:膜电容的变化可能来自线性和非线性源。例如,膜表面积或介电特性的变化可以线性地改变电容,而电压依赖性蛋白质的传感器残基可以非线性地改变电容。在这里,我们检查了红外线(IR)激光在全细胞电压钳控制下的对照和Prestin(SLC26a5)转染的人胚肾(HEK)细胞中引起的温度快速跳跃的影响。 Prestin的电压传感器具有独特的钟形,电压依赖性非线性电容(NLC)。由于双层效应,不管保持电压如何,控制型HEK电池中的温度跳跃都会导致膜电容(Cm)单相增加。然而,经Prestin转染的HEK细胞在Cm上还表现出双相增加/减少,其反向电位对应于Prestin的峰值NLC(Vh)处的电压,这归因于Vh的快速温度跟随漂移,其漂移速率高达在5毫秒的IR脉冲过程中为14 V / s。用水杨酸酯(一种已知的NLC抑制剂)进行治疗,可以重新建立控制细胞的行为。一个简单的动力学模型概括了我们的生物物理观察。这些结果证明了电压依赖性蛋白能够以双层敏感性对同等温度下的快速温度扰动做出反应。这可能是由于prestin独特的以千赫兹速率移动传感器电荷的能力,这是外毛细胞作为耳蜗放大器所必需的。

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