Applications of Genetic Code Expansion in Studying Protein Post-translational Modification

Area of Science:

• Biochemistry
• Molecular Biology
• Proteomics

Background:

• Protein post-translational modifications (PTMs) regulate diverse cellular functions but are challenging to study due to their dynamic nature.
• Site-specific PTMs are crucial for protein activity, localization, interaction, and stability.
• Understanding PTMs requires precise experimental tools to overcome challenges in deciphering their biological implications.

Purpose of the Study:

• To review the application of genetic code expansion for studying site-specific protein PTMs.
• To explore methods for incorporating PTMs or their mimics using non-canonical amino acids.
• To discuss the potential of genetic code expansion in advancing PTM research.

Main Methods:

• Genetic code expansion using orthogonal aminoacyl-tRNA synthetase/tRNA pairs to incorporate non-canonical amino acids.
• Direct incorporation of non-canonical amino acids bearing PTMs or their mimics.
• Utilizing precursor non-canonical amino acids followed by chemical/enzymatic modifications.
• Genetically encoding amino acids for functional regulation of PTM-modifying enzymes.

Main Results:

• Demonstrated applicability for various PTMs including phosphorylation, sulfation, nitration, acetylation, methylation, and citrullination.
• Enabled study of acylation, ubiquitination, and other PTMs via precursor amino acids.
• Showcased potential for functional studies of PTM-related enzymes.

Conclusions:

• Genetic code expansion is a powerful tool for site-specific PTM research in vitro and in vivo.
• This technology provides new avenues for investigating the physiological roles of PTMs.
• Future perspectives include overcoming limitations and expanding applications in proteomics.