Quantifying the Oligomeric States of Membrane Article Proteins in Cells through Super-Resolution Localizations
Quantifying the Oligomeric States of Membrane Article Proteins in Cells through Super-Resolution Localizations
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DOI：
10.1021/acs.jpcb.8b10402
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发表时间：
2018-11-22
影响因子：
3.3
通讯作者：
Chen，Tai-Yen
中科院分区：
文献类型：
--
作者：
Xie，Xihong;Cheng，Yu-Shan;Chen，Tai-Yen
Transitions between different oligomeric states of membrane proteins are essential for proper cellular functions. However, the quantification of their oligomeric states in cells is technically challenging. Here we developed a new method to quantify oligomeric state(s) of highly expressed membrane proteins using the probability density function of molecule density (PDFMD) calculated from super-resolution localizations. We provided the theoretical model ofPDFMD, discussed the effects of protein concentration, cell geometry, and photophysics of fluorescent proteins onPDFMD, and provided experimental criteria for proper quantification of oligomeric states. This method was further validated using simulated single-molecule fluorescent movies and applied to two membrane proteins, UhpT and SbmA inE. coli. The study shows thatPDFMDis useful in quantifying oligomeric states of membrane proteins in cells that can help in understanding cellular tasks. Potential applications to proteins with higher oligomeric states under high concentration and limitations of our methodology were also discussed.
