期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 54, 期 8, 页码 2548-2551出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201409847
关键词
biophysics; biosensors; excluded-volume effect; FRET; macromolecular crowding
资金
- Ministry of Innovation, Science and Research of the State of North Rhine-Westphalia (Ruckkehrer-programm)
- Cluster of Excellence RESOLV - German Research Foundation (DFG) [EXC 1069]
- International Graduate School of Neuroscience (Ruhr-University Bochum (Germany))
Biomolecules evolve and function in densely crowded and highly heterogeneous cellular environments. Such conditions are often mimicked in the test tube by the addition of artificial macromolecular crowding agents. Still, it is unclear if such cosolutes indeed reflect the physicochemical properties of the cellular environment as the in-cell crowding effect has not yet been quantified. We have developed a macromolecular crowding sensor based on a FRET-labeled polymer to probe the macromolecular crowding effect inside single living cells. Surprisingly, we find that excluded-volume effects, although observed in the presence of artificial crowding agents, do not lead to a compression of the sensor in the cell. The average conformation of the sensor is similar to that in aqueous buffer solution and cell lysate. However, the in-cell crowding effect is distributed heterogeneously and changes significantly upon cell stress. We present a tool to systematically study the in-cell crowding effect as a modulator of biomolecular reactions.
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