| Autor: |
Ko T; Department of Chemistry, University of Pennsylvania, 231 South 34th Street Philadelphia, PA 19104-6323, USA.; Department of Chemistry, New York University, 100 Washington Square East, New York, NY 10003, USA., Jou C; Department of Psychology, Hunter College, 695 Park Avenue, New York, NY, 10065, USA., Grau-Perales AB; Center for Neural Science, New York University, 4 Washington Place, New York, NY 10003, USA., Reynders M; Department of Chemistry, New York University, 100 Washington Square East, New York, NY 10003, USA., Fenton AA; Center for Neural Science, New York University, 4 Washington Place, New York, NY 10003, USA., Trauner D; Department of Chemistry, University of Pennsylvania, 231 South 34th Street Philadelphia, PA 19104-6323, USA.; Department of Chemistry, New York University, 100 Washington Square East, New York, NY 10003, USA. |
| Abstrakt: |
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. |
