Selective N-Terminal Modification of Peptides and Proteins Using Fatty Acyl Phosphates

Area of Science:

• Chemical Biology
• Bioconjugation Chemistry
• Biotechnology

Background:

• Selective modification of proteins and peptides is crucial for biopharmaceutical production and studying post-translational modifications.
• Achieving selective N-terminal acylation over side chains in peptides and proteins remains a significant challenge.
• Existing methods often suffer from limited selectivity and narrow substrate scope.

Purpose of the Study:

• To develop a novel, highly selective method for N-terminal acylation of peptides and proteins.
• To address the limitations of current acylation techniques in chemical biology.
• To establish a versatile and bioorthogonal strategy for protein and peptide modification.

Main Methods:

• A biomimetic approach utilizing in situ enzymatic reagent activation (ERA) of carboxylic acids with ATP.
• Generation of acyl-adenosine monophosphates as reactive intermediates.
• Application of the ERA method to various peptides, proteins, and antibodies.

Main Results:

• The ERA method demonstrated high selectivity for the N-termini of peptides and proteins, including liraglutide, glucagon, and insulin.
• The acylation process tolerated a wide range of fatty acids, including azido and dicarboxylic acids, enabling bioorthogonal labeling.
• Successful application to antibody modification highlights the strategy's versatility.

Conclusions:

• The enzymatic reagent activation (ERA) acylation is a versatile and bioorthogonal method for selective N-terminal modification.
• This approach overcomes limitations of existing methods, offering broad applicability in bioconjugation.
• The developed strategy holds promise for producing stable peptide and protein conjugates and advancing biopharmaceutical development.