Multifunctional activity-based chemical probes for sirtuins

Area of Science:

• Biochemistry
• Chemical Biology
• Enzymology

Background:

• Sirtuins, NAD+-dependent protein deacylases, are crucial for numerous cellular processes.
• Developing chemical tools to study sirtuin activity is essential for functional annotation and drug discovery.

Purpose of the Study:

• To design and synthesize novel activity-based chemical probes (ABPs) for interrogating active sirtuins.
• To utilize these probes for visualizing, capturing, and perturbing sirtuin function in biological systems.

Main Methods:

• Development of ABPs with a tripeptide backbone, thioacyl warhead, photoaffinity group, and bioorthogonal tag.
• Application of probes in comparative studies to assess sirtuin levels under different conditions.
• Utilizing click chemistry for reporter conjugation (fluorophore, biotin) and PROTAC development.

Main Results:

• Probes successfully interrogated the functional state of active sirtuins in complex samples.
• Comparative analysis revealed changes in active sirtuin content under varying physiological conditions.
• Probes facilitated inhibitor discovery and enabled visualization and affinity capture of sirtuins.
• Developed PROTACs demonstrated effective degradation of human SIRT2.

Conclusions:

• The developed small molecule probes provide powerful tools for investigating sirtuin biological functions and physiological relevance.
• These probes offer new avenues for functional annotation and pharmacological modulation of sirtuins.
• The methodology enables advanced studies in enzymology and chemical biology related to sirtuins.