Enhanced detection in hydroxylamine probe for the quantitative and qualitative analysis of S-acylated fatty acids on proteins

Identifier:  TFG:9027

Handle: https://hdl.handle.net/20.500.11797/TFG9027

Authors:  Villaverde Nogues, Jahaciel

Abstract:
S-acylation is a reversible lipid modification that regulates protein localization, stability, and function, with key roles in signaling and disease. However, its study is limited by the low ionization efficiency of fatty acid hydroxamates (FAHs), released upon hydroxylamine cleavage. To improve detection sensitivity, FAHs were derivatized with pyridine-2-carbonyl chloride and 4,4-dimethoxypiperidine, enhancing their signal in HPLC-HRMS. This optimized method enabled robust quantitative and qualitative analysis of S-acylation, as demonstrated in the human Neuro-2a (N2a) cell line. This approach provides a powerful tool for investigating the heterogeneous S-acylome and its dysregulation in neurodegenerative diseases like neuronal ceroid lipofuscinosis (CLN1).