PROTEIN AGGREGATION DETECTION USING BODIPY-BASED FLUORESCENT PROBES
Open Access
Author:
Santoro, Leeann
Graduate Program:
Chemistry
Degree:
Master of Science
Document Type:
Master Thesis
Date of Defense:
February 06, 2019
Committee Members:
Xin Zhang, Thesis Advisor/Co-Advisor Philip C Bevilacqua, Committee Member Scott A Showalter, Committee Member
Keywords:
protein aggregate fluorophore Alzheimer's neurodegeneration fluorescence molecular rotor Huntington's ALS
Abstract:
Proteinaceous inclusions are a hallmark of numerous neurodegenerative diseases such as Alzheimer’s, Parkinson’s, amyotrophic lateral sclerosis (ALS), and Huntingtin’s disease. It is still unclear if the observed insoluble aggregates are the cause of toxicity or if they are simply a byproduct. There is growing evidence to suggest that the initial soluble clusters of protein aggregates (aka oligomers) are more toxic to cells than insoluble aggregates. Current methods do not permit visualization of oligomers in vivo, making it difficult to understand their behavior in relevant biological contexts. In this work, we developed an aggregation-detection probe using BODIPY: a viscosity-sensitive fluorescent molecular rotor (FMR). By exploiting the increased rigidity within protein aggregates, we have constructed a fluorescent probe that reports the presence of soluble oligomers of a given protein-of-interest via “turn-on” fluorescence. This thesis research provides a novel biochemical method with which to investigate previously invisible soluble oligomers and is the first application of BODIPY to detect and visualize protein aggregation in live cells.
