期刊
ANALYTICAL CHEMISTRY
卷 88, 期 22, 页码 10860-10866出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b00032
关键词
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资金
- National Natural Science Foundation of China [21475121]
- Fundamental Research Funds for the Central Universities [WK6030000026, WK3460000002]
- Recruitment Program of Global Expert
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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