Bioorthogonal pro-metabolites for profiling short chain fatty acylation

Area of Science:

• Biochemistry
• Cell Biology
• Metabolomics

Background:

• Short chain fatty acids (SCFAs) are crucial signaling molecules involved in health and disease.
• Short chain fatty acylation, a posttranslational modification (PTM), links metabolism to protein function.
• Existing methods for studying short chain fatty acylation are limited.

Purpose of the Study:

• To develop a novel bioorthogonal pro-metabolite strategy for profiling short chain fatty acylation in living cells.
• To discover new protein targets of SCFAs and understand their role in cellular processes.
• To provide new tools for studying SCFA signaling in differentiation and disease.

Main Methods:

• Synthesis of ester-caged bioorthogonal short chain fatty acids inspired by tributyrin.
• Cellular evaluation of synthesized pro-metabolites to identify effective probes.
• Comprehensive characterization of metabolic dependence, toxicity, and HDAC inhibitor sensitivity.
• Application of optimized probes to identify novel protein targets of SCFAs.

Main Results:

• Discovery of an azido-ester pro-metabolite metabolized to acyl-coenzyme A (CoA).
• Demonstration of robust protein labeling profiles with the developed probes.
• Identification of novel candidate protein targets of SCFAs in cells.
• Characterization of the bioorthogonal pro-metabolites' properties and sensitivities.

Conclusions:

• The bioorthogonal pro-metabolite strategy offers an unbiased approach for profiling cellular protein acylation.
• This method advances the study of SCFA signaling pathways.
• The developed tools have potential applications in understanding differentiation and disease mechanisms.