Description
- Abstract:
- The ability to detect and adapt to changing conditions and circumstances is a requirement of life at both the organismal and cellular levels. Inside the cell, rapid and fine-tuned responses to environmental cues can be achieved through dynamic post-translational modifications (PTMs) of proteins. Significant insights into the activity and dependencies of signaling pathways can therefore be gleaned from global surveys of PTM events, and in particular protein phosphorylation, as it is absolutely essential in the signaling cascades mediating the downstream functions of receptor tyrosine kinase systems. In T cells, coordinated phosphorylation of tyrosine residues is critical for transmission of signals from the activated T cell receptor. Investigations into the dynamic phosphoproteome of T cells have been greatly assisted by advances in mass spectrometry, which allows for the identification and quantitation of thousands of PTMs simultaneously without a requirement for site-specific antibodies. However, despite the increasing resolution and decreasing cost of mass spectrometry assays, the complex network of phosphorylation events governing T cell activation has been only incompletely characterized. In particular, while it is now appreciated that numerous feedback pathways within the T cell phosphoproteome work together to regulate the initiation of immune responses, when and where feedback loops occur in the pathway is poorly understood. To better understand the role that critical T cell signaling components play in feedback regulation of the T cell receptor pathway, we conducted a phosphoproteomic study into the role of Phospholipase C-γ1 in mediating the activation state of the receptor-proximal signaling machinery. Additionally, we investigated the application of optimized chromatography configurations to T cell phosphoproteomics, and demonstrated the immense potential to achieve greater qualitative and quantitative insights into the T cell signaling machinery. Finally, to support the visualization, management, annotation, and analysis of this proteomic data, we designed and implemented new tools to expedite discoveries from large LC-MS datasets. The subject of this work is therefore the insights we have gained into the mechanisms governing T cell activation and regulation as well as the technical and technological innovations that made those insights possible.
- Notes:
- Thesis (Ph. D.)--Brown University, 2017
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Citation
Belmont, Judson,
"Technologies for Phosphoproteomic Interrogation of T Cell Signaling"
(2017).
Molecular Biology, Cell Biology, and Biochemistry Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://doi.org/10.26300/bhn2-nb74
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Molecular Biology, Cell Biology, and Biochemistry Theses and Dissertations
Theses and Dissertations for the Molecular Biology, Cell Biology, and Biochemistry department....