Journal
ANALYTICAL CHEMISTRY
Volume 88, Issue 22, Pages 10860-10866Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b00032
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Funding
- National Natural Science Foundation of China [21475121]
- Fundamental Research Funds for the Central Universities [WK6030000026, WK3460000002]
- Recruitment Program of Global Expert
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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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