Synthetic Rewiring of Virus-Like Particles via Circular Permutation Enables Modular Peptide Display and Protein...

Area of Science:

• Biotechnology
• Structural Biology
• Nanotechnology

Background:

• Virus-like particles (VLPs) are versatile protein-based nanoparticles with applications in drug delivery and vaccine development.
• Engineering VLPs for novel functions often involves modifying protein subunits, but natural evolutionary strategies like circular permutation remain underexplored.

Purpose of the Study:

• To investigate the utility of circular permutation for engineering self-assembling protein nanoparticles.
• To create and characterize a comprehensive circular permutation library of the MS2 coat protein.
• To demonstrate novel functionalities enabled by circular permutation in VLPs.

Main Methods:

• Construction of a circular permutation library for the MS2 coat protein dimer.
• Validation of new terminal locations using cryo-electron microscopy.
• Development of C-terminal peptide tagging and covalent protein encapsulation strategies.

Main Results:

• Identification of novel terminal locations in the MS2 coat protein through circular permutation.
• Successful C-terminal peptide tagging and stable protein encapsulation via covalent bonding.
• Demonstration that circular permutation expands VLP engineering potential beyond traditional methods.

Conclusions:

• Circular permutation is a powerful strategy for engineering self-assembling protein nanoparticles.
• This technique allows for the creation of VLPs with novel functionalities not possible with native proteins.
• The study provides a systematic framework for exploring VLP structural determinants and engineering new features.