RefSOFI for Mapping Nanoscale Organization of Protein-Protein Interactions in Living Cells.
RefSOFI for Mapping Nanoscale Organization of Protein-Protein Interactions in Living Cells.
复制标题
DOI：
10.1016/j.celrep.2015.12.036
复制
发表时间：
2016-01-12
期刊：
影响因子：
8.8
通讯作者：
Zhang J
中科院分区：
文献类型：
--
作者：
Hertel F;Mo GC;Duwé S;Dedecker P;Zhang J
It has become increasingly clear that protein-protein interactions (PPIs) are compartmentalized in nanoscale domains that define the biochemical architecture of the cell. Despite tremendous advances in super-resolution imaging, strategies to observe PPIs at sufficient resolution to discern their organization are just emerging. Here we describe a strategy in which PPIs induce reconstitution of fluorescent proteins (FPs) that are capable of exhibiting single-molecule fluctuations suitable for Stochastic Optical Fluctuation Imaging (SOFI). Subsequently, spatial maps of these interactions can be resolved in super-resolution in living cells. Using this strategy, termed reconstituted fluorescence-based SOFI (refSOFI), we investigated the interaction between the endoplasmic reticulum Ca2+ sensor STIM1 and the pore-forming channel subunit ORAI1, a crucial process in store-operated Ca2+ entry (SOCE). Stimulating SOCE does not appear to change the size of existing STIM1/ORAI1 interaction puncta at the ER-plasma membrane junctions, but results in an apparent increase in the number of interaction puncta.
