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General description:
One of the interests of the group lies in understanding the role of protein monoubiquitination in the regulation of several cellular pathways such as endocytosis and cell signalling.
The goal of my current research project is the identification of the specific proteins that become monoubiquitinated after different treatments in mammalian cells. I will address this issue using a proteomic approach.
Role in RUBICON:
I am treating a stable cell line expressing double-tagged ubiquitin with different stimuli and I am looking for proteins that are specifically monoubiquitinated in response to the treatment.
The purification protocol relies on a) in vitro proteasome digestion of cellular lysates, b) double affinity purification, c) reversed phase chromatography and LC-MS/MS analysis.
In vitro 26S proteasome treatment is performed to enrich the cell lysate in monoubiquitinated proteins that will be subsequently purified by affinity purification.
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Simona Polo's group
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