Deciphering protein long-chain S-acylation using mass spectrometry proteomics strategies

Area of Science:

• Biochemistry
• Proteomics
• Post-translational Modifications

Background:

• Protein long-chain S-acylation involves attaching fatty acids to cysteine residues via thioester bonds.
• This modification regulates protein localization, trafficking, and stability but is challenging to study due to its hydrophobic and labile nature.
• Previous proteomic workflows were complicated by the lipid modification, hindering research.

Purpose of the Study:

• To review recent advances in mass spectrometry-based strategies for studying protein S-acylation.
• To highlight key developments in the past decade for analyzing the S-acylome.
• To discuss emerging challenges and future directions in the field.

Main Methods:

• Direct and indirect mass spectrometry-based strategies are summarized.
• Key techniques include acyl-biotin exchange and lipid metabolic labeling.
• Novel enrichment and fragmentation methods are also discussed.

Main Results:

• Recent advances have significantly improved the sensitivity, site-specificity, and quantitation of S-acylation analysis.
• Mass spectrometry-based workflows now offer expanded capabilities for studying this modification.
• The review consolidates progress in understanding S-acylation over the last decade.

Conclusions:

• Despite challenges, mass spectrometry has revolutionized the study of protein S-acylation.
• Future research should focus on distinguishing lipid-specific modifications and improving quantification.
• Further exploration of the S-acylome holds potential for functional and therapeutic insights.