Inherently Emissive Puromycin Analogues for Live Cell Labelling

Area of Science:

• Medicinal Chemistry
• Molecular Biology
• Chemical Biology

Background:

• Puromycin is a natural antibiotic that inhibits protein synthesis by causing premature chain termination.
• Developing synthetic analogues with improved properties or novel functionalities is an active area of research.

Purpose of the Study:

• To synthesize and characterize novel puromycin derivatives incorporating an emissive thieno[3,4-d]-pyrimidine core.
• To evaluate the biological activity (translation inhibition, bactericidal effects) of these analogues.
• To assess the potential of these derivatives as fluorescent probes for newly synthesized peptides.

Main Methods:

• Synthesis of puromycin analogues with azetidine and 3,3-difluoroazetidine as dimethylamine surrogates.
• In vitro assays to determine translation inhibition and bactericidal activity.
• Cellular studies using HEK293T cells and rat hippocampal neurons to visualize peptide labeling.

Main Results:

• Puromycin derivatives demonstrated translation inhibition and bactericidal activity comparable to the natural antibiotic.
• The analogues enabled cellular puromycylation, producing emissive products directly.
• The 3,3-difluoroazetidine analogue effectively fluorescently labeled newly translated peptides in live and fixed cells.

Conclusions:

• Novel emissive puromycin analogues were successfully synthesized and exhibit potent biological activity.
• These compounds serve as effective fluorescent probes for real-time visualization of protein synthesis.
• The developed analogues offer a valuable tool for studying translation dynamics in biological systems.