Abstract
Protein interactions underlie the complexity of neuronal function. Potential interactions between specific proteins in the brain are predicted from assays based on genetic interaction and/or biochemistry. Genetic interaction reveals endogenous, but not necessarily direct, interactions between the proteins. Biochemistry-based assays, on the other hand, demonstrate direct interactions between proteins, but often outside their native environment or without a subcellular context. We aimed to achieve the best of both approaches by visualizing protein interaction directly within the brain of a live animal. Here, we show a proof-of-principle experiment in which the Cdc42 GTPase associates with its alleged partner WASp within neurons during the time and space that coincide with the newly developing CNS.
Highlights
Protein interactions are physical events that take place at nanometer scales
From Molecules to Neurons to a Brain Networks evolve because their links change dynamically over time and space
We showed changes in a specific link in the protein-protein interaction network within the developing brain through a method that offers spatial and temporal resolution
Summary
Protein interactions are physical events that take place at nanometer scales. Dissociation constants indicate that noncovalent bonds that form between the interacting proteins can have a million-fold affinity advantage over casual encounters. The question of when and where a particular pair of interacting proteins engages in physical association is rarely investigated on the same experimental platform as the question of how it might contribute to synaptogenesis or any other aspects of neuronal differentiation. To address this challenge, we sought to visualize protein interactions directly in their native environment by utilizing Forster resonance energy transfer (FRET) [4]. By combining transgenics [5,6] with an imaging technology [7,8], we quantitate protein interactions by Cdc (cell division control protein 42 homolog) and its alleged signaling partner WASp (Wiskott–Aldrich Syndrome protein) within the native environment of a developing brain
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