Oxidation and Ubiquitination of synaptic proteins in aging and disease

Today, Alzheimer’s disease is one of the most important age-related neurodegenerative diseases, but its etiology remains still unknown. In neurodegenerative diseases, the ubiquitination is perturbed and may play a major role in the formation of hallmark structures such as senile plaques and tangles. Selective protein oxidation or mutations may cause protein damage and induce a proteasome dysfunction. Main objective of our group is the identification of key players in oxidation and ubiquitination, their role in aging and in diseases like Alzheimer’s disease, or in Schizophrenia with altered glutathione synthesis. For this human autopsy tissue is used, from frontal and entorhinal cortex of Alzheimer’s and control’s brains, to identify specific targets and to determine the influence of proteasome dysfunction and oxidative stress on synaptic proteins.

Our main goals are:

1.Quantification of oxidation and ubiquitination levels of synaptic and metabolic proteins

2.Determine if there is an influence of oxidative stress on synaptic proteins in aging and disease

3.Verify if there is a correlation between oxidation level and the alteration of some mitochondrial proteins

4.Determine if there is an alteration of proteasome system in Alzheimer’s brains compared to control ones

5.Verify if there is correlation between synaptic proteins alteration and the hallmark of Alzheimer’s disease the neurofibrillary tangles and senile plaques

For this, different methods will be used such as isolation of proteins by immunoprecipitation or via ubiQapture, 1D and 2D gel electrophoresis, MALDI-TOF, Western blots, immunohistochemistry with specific antibodies against oxidized and ubiquitinated proteins and confocal Microscopy.