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CHOLESTEROL ENRICHMENT ALTERS COMPARTMENTALIZATION OF G PROTEINS INTO LIPID RAFT MICRODOMAINS

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title
CHOLESTEROL ENRICHMENT ALTERS COMPARTMENTALIZATION OF G PROTEINS INTO LIPID RAFT MICRODOMAINS
author
Mayer, Jonathan Caleb
abstract
G protein-coupled receptors (GPCRs) are located within the plasma membrane, particularly cholesterol-enriched domains known as lipid rafts. The localization of G-proteins and GPCRs are modulated by cholesterol, and cholesterol depletion results in GPCR dysfunction. However, few studies have investigated the impact of increased cholesterol content on the localization of G-protein subunits. Using neuroblastoma 2A cells, this project examined the effects of cholesterol enrichment on the localization of various G-protein subunits. Discontinuous sucrose density gradient ultracentrifugation was utilized to fractionate cell membranes into lipid raft and non-raft regions following treatment with varying concentrations of cholesterol. We found that all G-protein subunits translocated into lipid raft microdomains. To further study the functional impact of G-protein subunit translocation, we determined the function of metabotropic glutamate receptor 2 (mGluR2) which couples to Gi/o following cholesterol addition. A cAMP detection assay measured a decrease in the inhibition of adenylyl cyclase upon mGluR2 agonist activation. We found that cholesterol enrichment reduced the ability of the mGluR2 agonist to inhibit cAMP production, although the localization of mGluR2 in lipid raft and non-raft microdomains was not altered. Our study provides an important insight into understanding GPCR dysregulation in neurological diseases accompanied by altered cholesterol content in the brain.
subject
Cholesterol
G Protein
GPCR
mGluR2
contributor
Chen, Rong (advisor)
Cervera-Juanes, Rita (committee member)
Howlett, Allyn (committee member)
date
2024-05-23T08:36:26Z (accessioned)
2024 (issued)
degree
Biomedical Science – MS (discipline)
embargo
2029-05-18 (terms)
2029-05-18 (liftdate)
identifier
http://hdl.handle.net/10339/109455 (uri)
language
en (iso)
publisher
Wake Forest University
type
Thesis

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