溶质转运体26A家族在HEK-293细胞中的表达和功能

Expression and functional assessment of solute carrier 26A transporter family in HEK-293 cells

目的构建能在HEK-293细胞膜上表达的溶质转运体26A家族(SLC26A)的蛋白质粒,并对其生理功能进行检测。方法构建多种人类SLC26A转运体-绿色荧光蛋白的融合质粒,并使用瞬时转染技术将其表达在人胚肾293细胞(HEK-293)中。用全细胞膜片钳技术记录表达有SLC26A蛋白的HEK-293细胞的膜电容。结果每个SLC26A转运体均能在HEK-293细胞的细胞膜上表达,并表现出和离子相互作用的生理功能。SLC26A转运体与离子结合的数目与其在细胞膜上的表达程度正相关并具有一定的离子选择性。结论 SLC26A转运体家族能够在HEK-293细胞上表达并具有与转运离子结合的生理功能。在HEK-293细胞系中表达SLC26A转运体家族的研究方法,为进一步研究此家族成员转运功能及其转运机制提供了可能。

Objective To express solute carrier 26 A proteins in HEK- 293 cells and explore their functions. Methods SLC26 Ae GFP plasmids were transiently transfected into HEK-293 cells, and the nonlinear capacitance of the cells expressing SLC26 A proteins was measured by whole-cell patch recording. Results All the SLC26 A transporters were expressed on the membrane of HEK- 293 cells. Each member of the SLC26 A transporter family showed robust nonlinear capacitance, which represented their binding capability with anions. Conclusion The SLC26 A transporters expressed on HEK cells show similar functions as expected in tissue environment. The plasmids we constructed facilitate structural and functional study of SLC26 A transporters.