Cargando…

A Photoactivated Protein Degrader for Optical Control of Synaptic Function

Hundreds of proteins determine the function of synapses, and synapses define the neuronal circuits that subserve myriad brain, cognitive, and behavioral functions. It is thus necessary to precisely manipulate specific proteins at specific sub-cellular locations and times to elucidate the roles of pa...

Descripción completa

Detalles Bibliográficos
Autores principales: Ko, T., Jou, C., Grau-Perales, A.B., Reynders, M., Fenton, A.A., Trauner, D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949324/
https://www.ncbi.nlm.nih.gov/pubmed/36824807
http://dx.doi.org/10.1101/2023.02.13.528397
_version_ 1784892952540086272
author Ko, T.
Jou, C.
Grau-Perales, A.B.
Reynders, M.
Fenton, A.A.
Trauner, D.
author_facet Ko, T.
Jou, C.
Grau-Perales, A.B.
Reynders, M.
Fenton, A.A.
Trauner, D.
author_sort Ko, T.
collection PubMed
description Hundreds of proteins determine the function of synapses, and synapses define the neuronal circuits that subserve myriad brain, cognitive, and behavioral functions. It is thus necessary to precisely manipulate specific proteins at specific sub-cellular locations and times to elucidate the roles of particular proteins and synapses in brain function. We developed PHOtochemically TArgeting Chimeras (PHOTACs) as a strategy to optically degrade specific proteins with high spatial and temporal precision. PHOTACs are small molecules that, upon wavelength-selective illumination, catalyze ubiquitylation and degradation of target proteins through endogenous proteasomes. Here we describe the design and chemical properties of a PHOTAC that targets Ca(2+)/calmodulin-dependent protein kinase II alpha (CaMKIIα), which is abundant and crucial for baseline synaptic function of excitatory neurons. We validate the PHOTAC strategy, showing that the CaMKIIα-PHOTAC is effective in mouse brain tissue. Light activation of CaMKIIα-PHOTAC removed CaMKIIα from regions of the mouse hippocampus only within 25 μm of the illuminated brain surface. The optically-controlled degradation decreases synaptic function within minutes of light activation, measured by the light-initiated attenuation of evoked field excitatory postsynaptic potential (fEPSP) responses to physiological stimulation. The PHOTACs methodology should be broadly applicable to other key proteins implicated in synaptic function, especially for evaluating their precise roles in the maintenance of long-term potentiation and memory within subcellular dendritic domains.
format Online
Article
Text
id pubmed-9949324
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Cold Spring Harbor Laboratory
record_format MEDLINE/PubMed
spelling pubmed-99493242023-02-24 A Photoactivated Protein Degrader for Optical Control of Synaptic Function Ko, T. Jou, C. Grau-Perales, A.B. Reynders, M. Fenton, A.A. Trauner, D. bioRxiv Article Hundreds of proteins determine the function of synapses, and synapses define the neuronal circuits that subserve myriad brain, cognitive, and behavioral functions. It is thus necessary to precisely manipulate specific proteins at specific sub-cellular locations and times to elucidate the roles of particular proteins and synapses in brain function. We developed PHOtochemically TArgeting Chimeras (PHOTACs) as a strategy to optically degrade specific proteins with high spatial and temporal precision. PHOTACs are small molecules that, upon wavelength-selective illumination, catalyze ubiquitylation and degradation of target proteins through endogenous proteasomes. Here we describe the design and chemical properties of a PHOTAC that targets Ca(2+)/calmodulin-dependent protein kinase II alpha (CaMKIIα), which is abundant and crucial for baseline synaptic function of excitatory neurons. We validate the PHOTAC strategy, showing that the CaMKIIα-PHOTAC is effective in mouse brain tissue. Light activation of CaMKIIα-PHOTAC removed CaMKIIα from regions of the mouse hippocampus only within 25 μm of the illuminated brain surface. The optically-controlled degradation decreases synaptic function within minutes of light activation, measured by the light-initiated attenuation of evoked field excitatory postsynaptic potential (fEPSP) responses to physiological stimulation. The PHOTACs methodology should be broadly applicable to other key proteins implicated in synaptic function, especially for evaluating their precise roles in the maintenance of long-term potentiation and memory within subcellular dendritic domains. Cold Spring Harbor Laboratory 2023-02-14 /pmc/articles/PMC9949324/ /pubmed/36824807 http://dx.doi.org/10.1101/2023.02.13.528397 Text en https://creativecommons.org/licenses/by-nc/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (https://creativecommons.org/licenses/by-nc/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Ko, T.
Jou, C.
Grau-Perales, A.B.
Reynders, M.
Fenton, A.A.
Trauner, D.
A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title_full A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title_fullStr A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title_full_unstemmed A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title_short A Photoactivated Protein Degrader for Optical Control of Synaptic Function
title_sort photoactivated protein degrader for optical control of synaptic function
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9949324/
https://www.ncbi.nlm.nih.gov/pubmed/36824807
http://dx.doi.org/10.1101/2023.02.13.528397
work_keys_str_mv AT kot aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT jouc aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT grauperalesab aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT reyndersm aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT fentonaa aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT traunerd aphotoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT kot photoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT jouc photoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT grauperalesab photoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT reyndersm photoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT fentonaa photoactivatedproteindegraderforopticalcontrolofsynapticfunction
AT traunerd photoactivatedproteindegraderforopticalcontrolofsynapticfunction