Screening of voltage-sensitive proteins using Voltage-Sensing Phosphatase, VSP

我们的目的是通过与电压敏感磷酸酶的嵌合蛋白制备，开发一种新型电压敏感膜蛋白模块的功能筛选系统。首先，我们确定了Ci-VSP是最适合开发系统的VSP同源物，因为PI(4,5)P2磷酸酶活性的读数最显著。膜电压对ci - vsp酶活性的增强更为显著。其他电压传感器融合到Ci-VSP的细胞质区域，获得嵌合电压感应磷酸酶。

We aimed to develop a functional screening system for novel voltage-sensitive membrane protein modules through making a chimeric protein with voltage-sensing phosphatase. First, we identified Ci-VSP as the most suitable among VSP orthologsfor developing the system, since readout of PI(4,5)P2 phosphatase activity was most remarkable. Ci-VSPwas remodeled into a form in which enzyme activity was more remarkablyenhanced by membrane voltage. Other voltage sensors were fused to the cytoplasmic region of Ci-VSP to acquire chimeric voltage sensing phosphatases.