Tau protein is a microtubule-associated protein that is mainly found in nerve cells and is involved in the stabilization of microtubules. In certain neurodegenerative diseases, such as Alzheimer's disease (Alzheimer).'S disease, AD), tau proteins may be abnormally phosphorylated and aggregated, forming so-called tau clumps or neurofibrillary tangles.
Ubiquitination is a post-translational modification of a protein that involves attaching one or more ubiquitin molecules to a target protein. This modification typically labels proteins for degradation, but is also involved in many other cellular processes.
Figure 1Ubiquitination modification research route.
The identification of ubiquitination of tau proteins is primarily to determine whether tau proteins are ubiquitinated and at which sites ubiquitination occurs. Here are the basic steps:
1. Sample preparation:
Tau protein is extracted from relevant cells or tissues.
2. Immunoprecipitation:
Immunoprecipitation was performed using antibodies against tau protein to enrich tau protein from the sample.
3. Protein electrophoresis:
The precipitated proteins were separated by SDS-PAGE.
4. Immunoblotting analysis:
Western blot analysis using antibodies against ubiquitin was performed to detect whether tau protein was ubiquitinated.
5. Mass spectrometry analysis
In order to accurately identify ubiquitination sites on tau proteins, further mass spectrometry analysis can be performed.
6. Data analysis
Resolve the ubiquitination sites of tau protein based on mass spectrometry data.
Ubiquitination of tau protein may be associated with its aggregation and degradation mechanisms in disease, so its identification is important to understand the molecular mechanisms of neurodegenerative diseases.