Design, synthesis and evaluation of chemical probes for biochemical applications
Date Issued
2015
Date
2015
Author(s)
Hsu, Yu-Ling
Abstract
Development of small-molecular tools which selectively react with designated protein families has been found powerful in modern functional proteomics. In this dissertation, we evaluated two kinase probe libraries according to the structure and functions of protein kinases. The first library adopted a 5''-p-fluorosulfonylbenzoyl adenosine (5''-FSBA) skeleton and a biotin reporter. The second library explored a novel molecular framework by attaching three kind of electrophilic warheads on 3''-azido 2'', 3''-dideoxy base (AZT and AZA) recognition unit. The azido group at 3'' position is a native clickable tag without further modification. The labeling performances of these two probe libraries toward kinases were compared. Despite the therapeutic importance of this multifunctional herbal compound Andrographolide, the inhibition and regulation mechanism are still ambiguious. Herein, we designed a novel andrographolide-based, cell permeable fluorescent probe for in vivo identification of target proteins participating in related cancer or diseases. This first-time synthesized Andrographolide-based, cell permeable fluorescent probe provide valuable information for the synthesis method in the future. In addition, cell-permeable activity-based probes for α-L-fucosidase were evaluated by the key base-promoted epimerization step. These two probes carrying mono or difluoromethylphenyl group would generate reactive quinone methide as a trapping device after hydrolysis. In the last part, quantitative isotope-coded azido tags were synthesized and applied to detect the level of S-nitrosylation proteome with drug treatment.
Subjects
probe
protein profiling
andrographolide
α-L-fucosidase
quinone methide
isotope-coded tag
SDGs
Type
thesis