化学
钙调蛋白
体外
细胞内
生物物理学
金属
钙
内生
生物化学
酶
生物
有机化学
作者
Gongyu Li,Siming Yuan,Yang Pan,Yangzhong Liu,Guangming Huang
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2016-10-17
卷期号:88 (22): 10860-10866
被引量:27
标识
DOI:10.1021/acs.analchem.6b00032
摘要
The identification of endogenous proteins as well as their binding to metal ions in living cells is determined by combining pulsed electrophoretic separations with nanoelectrospray ionization followed by mass spectrometric detection. This approach avoids problems resulting from the complicated cellular environment. In this manner, we demonstrate the rapid identification (300 ms or less) of intact proteins from living E. coli cells including the complexation of calmodulin with calcium ion. The latter showed different binding states from those observed in in vitro studies. These observations also reveal in vitro measurements do not necessarily represent the actual situation in living cells. We conclude that the attempted in situ measurement of intracellular proteins with minimal sampling processes should be preferred.
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