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Title: RefSOFI for Mapping Nanoscale Organization of Protein-Protein Interactions in Living Cells
Authors: Hertel, Fabian ×
Mo, Gary C. H
Duwé, Sam
Dedecker, Peter
Zhang, Jin #
Issue Date: 2016
Publisher: Elsevier Inc
Series Title: Cell Reports vol:14 issue:2 pages:390-400
Abstract: It has become increasingly clear that protein-protein interactions (PPIs) are compartmentalized in nano- scale domains that define the biochemical architec- ture of the cell. Despite tremendous advances in su- per-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 (ER) Ca2+ sensor STIM1 and the pore-forming chan- nel subunit ORAI1, a crucial process in store-oper- ated Ca2+ entry (SOCE). Stimulating SOCE does not appear to change the size of existing STIM1/ORAI1 interaction puncta at the ER-plasmamembrane junc- tions, but results in an apparent increase in the num- ber of interaction puncta.
ISSN: 2211-1247
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Molecular Imaging and Photonics
Biochemistry, Molecular and Structural Biology Section
× corresponding author
# (joint) last author

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