A split horseradish peroxidase for detection of intercellular protein-protein interactions and sensitive visualization of synapses

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A split horseradish peroxidase for detection of intercellular protein-protein interactions and sensitive visualization of synapses

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Title: A split horseradish peroxidase for detection of intercellular protein-protein interactions and sensitive visualization of synapses
Author: Martell, Jeffrey D.; Yamagata, Masahito; Deerinck, Thomas J.; Phan, Sébastien; Kwa, Carolyn G.; Ellisman, Mark H.; Sanes, Joshua R.; Ting, Alice Y.

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Citation: Martell, Jeffrey D., Masahito Yamagata, Thomas J. Deerinck, Sébastien Phan, Carolyn G. Kwa, Mark H. Ellisman, Joshua R. Sanes, and Alice Y. Ting. 2016. “A split horseradish peroxidase for detection of intercellular protein-protein interactions and sensitive visualization of synapses.” Nature biotechnology 34 (7): 774-780. doi:10.1038/nbt.3563. http://dx.doi.org/10.1038/nbt.3563.
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Abstract: 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 techniques1,2,3 have insufficient sensitivity and/or specificity. Here we report split horseradish peroxidase (sHRP) as a sensitive and specific tool for 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 cleft4, 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 for studying mechanisms of communication between a variety of cell types.
Published Version: doi:10.1038/nbt.3563
Other Sources: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4942342/pdf/
Terms of Use: This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA
Citable link to this page: http://nrs.harvard.edu/urn-3:HUL.InstRepos:29626184
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