Type
Text
Type
Thesis
Advisor
Bowen, Mark E | Miller, W. Todd
Date
2016-12-01
Keywords
NMDAR, Palmitoylation, Phosphonatetheinyl Transferase, PPTase, PSD95 | Biochemistry
Department
Department of Biochemistry and Cell Biology
Language
en_US
Source
This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.
Identifier
http://hdl.handle.net/11401/76886
Publisher
The Graduate School, Stony Brook University: Stony Brook, NY.
Format
application/pdf
Abstract
Signal transduction relies on the proper localization of proteins to properly transfer signals throughout the cell. The cell membrane serves as a vital interface for the proper localization of proteins. Posttranslational modifications cause proteins to become targeted to membranes. One such modification is palmitoylation, the addition of a palmitic acid to a cysteine of a protein. Within the neuron, a large protein complex called the postsynaptic density receives upstream signals at the synapse. Many signaling proteins within the postsynaptic density are known to require palmitoylation for proper trafficking and function. Two such examples are the N-methyl-D-Aspartate receptor (NMDAR) and Post Synaptic Density Protein 95 (PSD95). While studies have been carried out on their soluble forms, their palmitoylated forms have not been well studied. This is due to the difficulty in palmitoylating proteins in-vitro. We circumvent this issue by utilizing phosphopantetheinyl transferases (PPTases) to palmitoyate proteins. PPTases are able to attach various Coenzyme A substrates to proteins with specific tag sequences. We also develop a 2-step assay to quantify the effectiveness of the palmitoylation reaction. We successfully show that palmitoylation using AcpS, a PPTase, is possible utilizing the developed assays. | 47 pages
Recommended Citation
Chang, Victor, "Palmitoylation of the NMDA receptor and PSD95 utilizing Phosphopantetheinyl Transferases" (2016). Stony Brook Theses and Dissertations Collection, 2006-2020 (closed to submissions). 2760.
https://commons.library.stonybrook.edu/stony-brook-theses-and-dissertations-collection/2760