Enzyme-Directed Functionalization of Designed, Two-Dimensional Protein Lattices.

Area of Science:

• Biotechnology and Materials Science
• Protein Engineering and Nanotechnology

Background:

• Crystalline protein arrays offer potential in sensing, catalysis, and medicine.
• Generating artificial functional materials from protein assemblies remains underexplored.
• Enzyme-directed post-translational modifications are key to proteome diversity and could functionalize artificial assemblies.

Purpose of the Study:

• To explore the generation of artificial functional materials using protein assemblies.
• To utilize enzyme-directed modifications for selective tailoring of protein crystal surfaces.
• To demonstrate chemoenzymatic modification of 2D protein crystals for advanced applications.

Main Methods:

• Employed phosphopantetheinyl transferases (PPTases) for site-selective protein modification.
• Used a peptide tag (ybbR) or molecular tag (CoA) for covalent tethering to 2D protein arrays.
• Utilized Sfp PPTase for enzymatic functionalization of the modified arrays.

Main Results:

• Achieved site-specific surface tailoring of designed 2D protein crystals.
• Demonstrated successful covalent tethering of peptide or CoA tags.
• Generated small molecule- and protein-functionalized surfaces without compromising crystalline order.

Conclusions:

• PPTases enable chemoenzymatic modification of large protein surfaces.
• This approach facilitates the creation of sophisticated protein platforms.
• The engineered platforms mimic the complexity of cell surfaces for diverse applications.