Gαi3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins

Thomas S. Weiss; Chester E. Chamberlain; Tetsuro Takeda

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Gαi3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins

机译：绿色荧光蛋白融合蛋白的荧光共振能量转移监测Gαi3与活细胞高尔基体膜上的钙蛋白的结合

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Gαi3 is found both on the plasma membrane and on Golgi mem- branes. Calnuc, an EF hand protein, binds both Gαi3 and Ca~2+ and is found both in the Golgi lumen and in the cytoplasm. To inves- tigate whether Gαi3 binds calnuc in living cells and where this interaction takes place we performed fluorescence resonance en- ergy transfer (FRET) analysis between Gαi3 and calnuc in COS-7 cells expressing Gαi3-yellow fluorescent protein (YFP) and calnuc- cyan fluorescent protein (CFP). The tagged proteins have the same localization as the endogenous, nontagged proteins. When Gαi3- YFP and calnuc-CFP are coexpressed, a FRET signal is detected in the Golgi region, but no FRET signal is detected on the plasma mem- brane.

机译：在质膜和高尔基膜上都发现了Gαi3。 Calnuc是一种EF手蛋白，与Gαi3和Ca〜2 +结合，并且在高尔基体腔和细胞质中均发现。为了研究Gαi3是否与活细胞中的钙蛋白结合以及在何处发生相互作用，我们在表达Gαi3黄色荧光蛋白（YFP）和钙蛋白的COS-7细胞中，对Gαi3和钙蛋白之间进行了荧光共振能量转移（FRET）分析。青色荧光蛋白（CFP）。标记的蛋白质与内源的非标记的蛋白质具有相同的定位。当Gαi3-YFP和calnuc-CFP共表达时，在高尔基体区域检测到FRET信号，但在血浆膜上未检测到FRET信号。

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《Proceedings of the National Academy of Sciences of the United States of America》 |2001年第26期|p.14961-14966|共6页
作者
Thomas S. Weiss; Chester E. Chamberlain; Tetsuro Takeda;
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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种 eng
中图分类自然科学总论;
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5. Detecting the Conformation of Individual Proteins in Live Cells using Single Molecule Fluorescence Resonance Energy Transfer [D] . Sakon, John Joseph 2010

机译：使用单分子荧光共振能量转移检测活细胞中单个蛋白质的构象
6. Gαi3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins [O] . Thomas S. Weiss, Chester E. Chamberlain, Tetsuro Takeda, 2001

机译：Gαi3与活细胞高尔基体膜上的钙蛋白结合通过绿色荧光共振能量转移进行监测荧光蛋白融合蛋白
7. Gαi3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins [O] . Weiss, Thomas S., Chamberlain, Chester E., Takeda, Tetsuro, 2001

机译：Gαi3与活细胞高尔基体膜上的钙蛋白结合通过绿色荧光共振能量转移进行监测荧光蛋白融合蛋白

Gαi3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins

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