Mass Spectrometry-Based Global Proteomic Analysis of Endoplasmic Reticulum and Mitochondria Contact Sites

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Overview

Title
Mass Spectrometry-Based Global Proteomic Analysis of Endoplasmic Reticulum and Mitochondria Contact Sites
Contributors
Poston, Chloe N (creator)
Bazemore-Walker, Carthene (Director)
Delaney, Sarah (Reader)
Suggs, John (Reader)
Brown University. Chemistry (sponsor)
Doi
10.7301/Z03N21Q4
Copyright Date
2013
Abstract
Mass spectrometry has been a long-standing analytical tool for chemists. Tandem mass spectrometry-based proteomics capitalizes on the sensitivity, accuracy, and efficiency of this classic technique in order to investigate the protein content of specific sub-cellular regions. In this work we apply tandem mass spectrometry (MS/MS) to the analysis of mitochondria-associated endoplasmic reticulum membranes (MAMs). MAMs are a point of communication between the ER and the mitochondria that facilitate Ca2+ trafficking, protein folding, and energy metabolism. They have been implicated in the regulation of apoptosis and more recently neurodegenerative disorders including Alzheimer's disease and Parkinson's. The involvement of the MAM in these diseases has yet to be determined, largely because proteins localized to the MAM have yet to be elucidated. We address this gap in knowledge with our tandem mass spectrometry-based global characterization of MAMs from mouse brain tissue. We employ a gel-based LC/LC-MS/MS method that allows for the analysis of hydrophobic membrane proteins. Our analysis reveals 1,212 unique proteins in the MAM that are involved in cell signaling, small molecule trafficking, and protein processing. Bio-informatic analysis of our identified MAM proteins supports postulates that the MAM may play a role in the pathology of Alzheimer's disease, schizophrenia, and dyskinesia. Our data also suggests that lipid rafts within the MAM are involved in signaling and transport within the region, and may play a role in the MAM's relationship to neurodegenerative disease. The completion of this work provides a much-anticipated catalog of proteins at the MAM, which can be used for targeted studies to elucidate pathways and novel therapeutic targets in the region.
Keywords
Mitochondria-Associated Membranes
Mass spectrometry
Proteomics
Notes
Thesis (Ph.D. -- Brown University (2013)
Extent
x, 154 p.

Citation

Poston, Chloe N., "Mass Spectrometry-Based Global Proteomic Analysis of Endoplasmic Reticulum and Mitochondria Contact Sites" (2013). Chemistry Theses and Dissertations. Brown Digital Repository. Brown University Library. https://doi.org/10.7301/Z03N21Q4

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