Global Profiling of Acetyltransferase Feedback Regulation

Area of Science:

• Biochemistry
• Cellular Signaling
• Proteomics

Background:

• Lysine acetyltransferases (KATs) are crucial enzymes regulating cell signaling pathways.
• Understanding KAT regulation is vital for deciphering cellular processes and disease mechanisms.
• Existing methods for profiling KAT interactions have limitations in scope and sensitivity.

Purpose of the Study:

• To develop and optimize a chemoproteomic platform for profiling KAT-ligand interactions in complex biological samples.
• To comprehensively analyze the inhibition of KATs by the metabolite coenzyme A (CoA).
• To identify novel KATs regulated by metabolic feedback.

Main Methods:

• Utilized inhibitor-functionalized capture resins for affinity-based enrichment of KATs and associated complexes.
• Applied chemoproteomic profiling to complex proteomes to identify KAT-ligand interactions.
• Conducted a comprehensive analysis of KAT inhibition by CoA.

Main Results:

• The optimized platform successfully profiled a broader range of KATs, KAT complexes, and CoA-dependent enzymes.
• Revealed a spectrum of sensitivity to CoA across the KAT superfamily.
• Identified Nucleosome Acetyltransferase of type 1 (NAT10) as a novel KAT susceptible to CoA-mediated feedback inhibition.

Conclusions:

• The developed chemoproteomic platform offers a powerful tool for studying KAT regulation.
• The findings provide new insights into the metabolic control of KAT-dependent signaling pathways.
• This approach can be used to assess the potency and selectivity of reversible regulators of KATs, including small molecules and metabolites.