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A split horseradish peroxidase for the detection of intercellular protein–protein interactions and sensitive visualization of synapses
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A split horseradish peroxidase for the detection of intercellular protein–protein interactions and sensitive visualization of synapses
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Journal Article
A split horseradish peroxidase for the detection of intercellular protein–protein interactions and sensitive visualization of synapses
2016
Overview
Synapses can be detected with high sensitivity by a split reporter that visualizes intercellular protein–protein interactions.
Intercellular protein–protein interactions (PPIs) enable communication between cells in diverse biological processes, including cell proliferation, immune responses, infection, and synaptic transmission, but they are challenging to visualize because existing techniques
1
,
2
,
3
have insufficient sensitivity and/or specificity. Here we report a split horseradish peroxidase (sHRP) as a sensitive and specific tool for the detection of intercellular PPIs. The two sHRP fragments, engineered through screening of 17 cut sites in HRP followed by directed evolution, reconstitute into an active form when driven together by an intercellular PPI, producing bright fluorescence or contrast for electron microscopy. Fusing the sHRP fragments to the proteins neurexin (NRX) and neuroligin (NLG), which bind each other across the synaptic cleft
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, enabled sensitive visualization of synapses between specific sets of neurons, including two classes of synapses in the mouse visual system. sHRP should be widely applicable to studying mechanisms of communication between a variety of cell types.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 13/109
/ 13/51
/ 14
/ 14/1
/ 14/19
/ 14/28
/ 14/34
/ 14/35
/ 14/63
/ 82
/ Animals
/ Biomedical Engineering/Biotechnology
/ Cloning
/ Enzymes
/ Horseradish Peroxidase - chemistry
/ Horseradish Peroxidase - genetics
/ Horseradish Peroxidase - pharmacokinetics
/ letter
/ Mice
/ Microscopy, Electron - methods
/ Microscopy, Fluorescence - methods
/ Nerve Tissue Proteins - metabolism
/ Protein Engineering - methods
/ Protein Interaction Mapping - methods
/ Protein-protein interactions
/ Proteins
/ Synapses
/ Yeast
