Voltage Clamp Fluorometry of P-Type ATPases.

Voltage Clamp Fluorometry of P-Type ATPases.
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P 型 AT 酶的电压钳荧光测定法。

DOI:
10.1007/978-1-4939-3179-8_25
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发表时间:
2016
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Dempski,RobertE
Dempski,RobertE
中科院分区:
--
文献类型:
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作者:
Dempski,RobertE

文献摘要

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电压钳荧光测定法已成为比较 P 型 ATP 酶(例如 Na+,K+-ATP 酶和 H+,K+-ATP 酶)的部分反应与这些离子泵的构象动力学的强大工具。在这里,我们描述了在 X 中异源表达膜蛋白的方法。左旋细胞并用一种或多种荧光团对这些蛋白质进行位点特异性标记。在两电极电压钳条件下,荧光变化与电流测量同时进行。
Voltage clamp fluorometry has become a powerful tool to compare partial reactions of P-type ATPases such as the Na+,K+-ATPase and H+,K+-ATPase with conformational dynamics of these ion pumps. Here, we describe the methodology to heterologously express membrane proteins inX. laevisoocytes and site-specifically label these proteins with one or more fluorophores. Fluorescence changes are measured simultaneously with current measurements under two-electrode voltage clamp conditions.