Quantifying Subcellular Ubiquitin-proteasome Activity in the Rodent Brain
| Autor: | Taylor McFadden, Rishi K Devulapalli, Timothy J. Jarome |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Male
Proteasome Endopeptidase Complex General Chemical Engineering Hippocampus Protein degradation medicine.disease_cause General Biochemistry Genetics and Molecular Biology Rats Sprague-Dawley Synapse 03 medical and health sciences 0302 clinical medicine Ubiquitin Protein targeting medicine Animals Cellular compartment 030304 developmental biology Cell Nucleus 0303 health sciences General Immunology and Microbiology biology General Neuroscience Ubiquitination Brain Amygdala Cell biology Proteasome Proteolysis Synapses Synaptic plasticity biology.protein 030217 neurology & neurosurgery |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/59695 |
| Popis: | The ubiquitin-proteasome system is a key regulator of protein degradation and a variety of other cellular processes in eukaryotes. In the brain, increases in ubiquitin-proteasome activity are critical for synaptic plasticity and memory formation and aberrant changes in this system are associated with a variety of neurological, neurodegenerative and psychiatric disorders. One of the issues in studying ubiquitin-proteasome functioning in the brain is that it is present in all cellular compartments, in which the protein targets, functional role and mechanisms of regulation can vary widely. As a result, the ability to directly compare brain ubiquitin protein targeting and proteasome catalytic activity in different subcellular compartments within the same animal is critical for fully understanding how the UPS contributes to synaptic plasticity, memory and disease. The method described here allows collection of nuclear, cytoplasmic and crude synaptic fractions from the same rodent (rat) brain, followed by simultaneous quantification of proteasome catalytic activity (indirectly, providing activity of the proteasome core only) and linkage-specific ubiquitin protein tagging. Thus, the method can be used to directly compare subcellular changes in ubiquitin-proteasome activity in different brain regions in the same animal during synaptic plasticity, memory formation and different disease states. This method can also be used to assess the subcellular distribution and function of other proteins within the same animal. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|