Investigating the turnover of N-acyl amino acids
Publication Date
July 31, 2026
Creator
Alrasheed, Arwa
Abstract
The metabolism of N-acyl amino acids (NAAs) is not fully understood, especially the enzymes responsible for their metabolism. The hypothesis examined in this thesis is that N-arachidonoyl glycine (NAGly) is metabolized by serine hydrolases, since it has a similar structure to anandamide, which is hydrolysed by the serine hydrolase fatty acid amide hydrolase (FAAH).This study aimed to investigate NAA metabolism using both a fluorometric assay and activity-based protein profiling (ABPP). A fluorometric assay was developed to detect amino acids, mainly glycine, released from NAA hydrolysis using derivatization reagents such as (Naphthalene-2,3-carboxaldehyde NDC) and fluoresceine. Several conditions were investigated, for example, reagent concentration, pH, and solvent. While fluoresceine improved sensitivity, the overall assay showed low levels of fluorescent product and high variability, limiting its effectiveness for detecting NAA hydrolysis. To address these problems, ABPP was used to identify active serine hydrolases in rat and human brain tissues. In human brain, several enzyme activities were detected, with some bands consistent with (Monoacylglycerol lipase )MAGL and FAAH based on molecular weight and inhibitor selectivity. In rat brain tissue, however, none of the enzyme activities were significantly inhibited using a wide range of serine hydrolase inhibitors. NAGly failed to alter the activity of any enzymes detected with this method. Overall, while ABPP is useful to detect multiple enzyme activities in complex proteomes, further refinements are needed for confident enzyme identification and determination of pathways of NAA turnover.
Item Type
ethesis
Thesis Type
Mphil
Supervisors
University of Nottingham
University of Nottingham
Subjects (LC)
Associated Schools / Departments
UoN Repository URI
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Mphil thesis corrections.pdf
Size
1.79 MB
Format
Adobe PDF
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