Designed Proteins as Novel Imaging Reagents in Living Escherichia coli

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 b...

Full description

Saved in:
Bibliographic Details
Published in:Chembiochem : a European journal of chemical biology 2016-09, Vol.17 (17), p.1652-1657
Main Authors: Pratt, Susan E., Speltz, Elizabeth B., Mochrie, Simon G. J., Regan, Lynne
Format: Article
Language:English
Subjects:
Bacterial Proteins - analysis
Bacterial Proteins - metabolism
Cytoskeletal Proteins - analysis
Cytoskeletal Proteins - metabolism
E coli
Escherichia coli
Escherichia coli - cytology
Escherichia coli - metabolism
fluorescence imaging
fluorescent probes
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Microbial Viability
Peptides - chemistry
Peptides - genetics
Peptides - metabolism
protein design
Proteins
repeat protein
tag-probe system
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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. TRAPping E. coli: We introduced a no‐ vel in vivo imaging strategy based on a set of designed tetratricopeptide repeat affinity proteins (TRAPs) that specifically and reversibly interact with a short peptide tag. It is fully genetically encodable and requires no exogenous substrates. We demonstrate its utility by imaging the E. coli protein FtsZ.
ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201600252