AI Article Synopsis
- Fluorescence imaging is showcased as an effective method to observe protein functions inside living cells using a new genetically encodable strategy.
- This method involves tetratricopeptide repeat affinity proteins (TRAPs) that interact with a special peptide tag, allowing for the addition of a small tag to proteins without disrupting their function.
- The research demonstrated its application using the protein FtsZ in E. coli, successfully tracking its structures in real-time under varying expression levels, indicating the strategy's potential for a wide range of proteins difficult to tag directly with fluorescent proteins.
Article Abstract
Fluorescence imaging is a powerful tool to study protein function in living cells. Here, we introduce a novel imaging strategy that is fully genetically encodable, does not require the use of exogenous substrates, and adds a minimally disruptive tag to the protein of interest (POI). Our method was based on a set of designed tetratricopeptide repeat affinity proteins (TRAPs) that specifically and reversibly interact with a short, extended peptide tag. We co-expressed the TRAPs fused to fluorescent proteins (FPs) and the peptide tags fused to the POIs. We illustrated the method using the Escherichia coli protein FtsZ and showed that our system could track distinct FtsZ structures under both low and high expression conditions in live cells. We anticipate that our imaging strategy will be a useful tool for imaging the subcellular localization of many proteins, especially those recalcitrant to imaging by direct tagging with FPs.
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| http://dx.doi.org/10.1002/cbic.201600252 | DOI Listing |
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