Thermostable exoshells fold and stabilize recombinant proteins

• Main Authors: Deshpande, Siddharth, Masurkar, Nihar D., Girish, Vallerinteavide Mavelli, Desai, Malan, Chakraborty, Goutam, Chan, Juliana M., Drum, Chester L.
• Other Authors: School of Chemical and Biomedical Engineering
• Format: Article
• Language: English
• Published: 2018
• Subjects: Nanoparticles; Protein Folding
• Online Access: https://hdl.handle.net/10356/89629; http://hdl.handle.net/10220/44987
• Institution: Nanyang Technological University

The expression and stabilization of recombinant proteins is fundamental to basic and applied biology. Here we have engineered a thermostable protein nanoparticle (tES) to improve both expression and stabilization of recombinant proteins using this technology. tES provides steric accommodation and charge complementation to green fluorescent protein (GFPuv), horseradish peroxidase (HRPc), and Renilla luciferase (rLuc), improving the yields of functional in vitro folding by ~100-fold. Encapsulated enzymes retain the ability to metabolize small-molecule substrates, presumably via four 4.5-nm pores present in the tES shell. GFPuv exhibits no spectral shifts in fluorescence compared to a nonencapsulated control. Thermolabile proteins internalized by tES are resistant to thermal, organic, chaotropic, and proteolytic denaturation and can be released from the tES assembly with mild pH titration followed by proteolysis.