Self-cyclisation as a general and efficient platform for peptide and protein macrocyclisation.

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Macrocyclisation of proteins and peptides significantly enhances structural stability.
• Cyclic peptides and proteins are valuable in drug discovery and as research tools (e.g., cyclised nanodiscs for membrane protein studies).
• Existing enzymatic methods for macrocyclisation have limitations.

Purpose of the Study:

• To engineer a self-cyclising protein, termed "autocyclase", for controllable, high-yield production of cyclic biomolecules.
• To elucidate the reaction mechanism of this unimolecular cyclisation process.
• To demonstrate the utility of autocyclases in generating diverse macrocyclic peptides and proteins.

Main Methods:

• Protein engineering to create a self-cyclising "autocyclase" enzyme.
• Characterisation of the autocyclisation reaction mechanism.
• Application of the autocyclase method to synthesise various cyclic peptides and proteins.

Main Results:

• Successful engineering of a self-cyclising autocyclase protein.
• Demonstration of a controllable, unimolecular reaction pathway for macrocyclisation.
• High-yield production of diverse cyclic peptides and proteins using the autocyclase system.
• Identification of alternative solutions to existing challenges in enzymatic cyclisation.

Conclusions:

• Autocyclases provide a simple and efficient alternative method for generating macrocyclic biomolecules.
• This engineered protein facilitates access to a wide range of cyclic peptides and proteins.
• The autocyclase approach has significant implications for drug discovery and biochemical research.