Protein Vesicles Self-Assembled from Functional Globular Proteins with Different Charge and Size.

Protein vesicles can be synthesized by mixing two fusion proteins: an elastin-like polypeptide (ELP) fused to an arginine-rich leucine zipper (Z) with a globular, soluble protein fused to a glutamate-rich leucine zipper (Z). Currently, only fluorescent proteins have been incorporated into vesicles; however, for protein vesicles to be useful for biocatalysis, drug delivery, or biosensing, vesicles must assemble from functional proteins that span an array of properties and functionalities. In this work, the globular protein was systematically changed to determine the effects of the surface charge and size on the self-assembly of protein vesicles. The formation of microphases, which included vesicles, coacervates, and hybrid structures, was monitored at different assembly conditions to determine the phase space for each globular protein. The results show that the protein surface charge has a small effect on vesicle self-assembly. However, increasing the size of the globular protein decreases the vesicle size and increases the stability at lower Z/Z molar ratios. The phase diagrams created can be used as guidelines to incorporate new functional proteins into vesicles. Furthermore, this work reports catalytically active enzyme vesicles, demonstrating the potential for the application of vesicles as biocatalysts or biosensors.